Fused cyclopentyl antagonists of ccr2

ABSTRACT

The present invention comprises compounds of Formula (I). 
     
       
         
         
             
             
         
       
     
     wherein: R 0 , R 1 , R 2 , R 3 , R 4 , R 5 , and A are as defined in the specification. The invention also comprises a method of preventing, treating or ameliorating a syndrome, disorder or disease, wherein said syndrome, disorder or disease is type II diabetes, obesity and asthma. The invention also comprises a method of inhibiting CCR2 activity in a mammal by administration of a therapeutically effective amount of at least one compound of Formula (I).

CROSS REFERENCE TO RELATED APPLICATION

This application claims priority from U.S. Provisional Application Ser.No. 61/621,138 filed Apr. 6, 2012, the contents of which are herebyincorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention is directed to substituted fused cyclopentyl compounds,which are antagonists to the chemoattractant cytokine receptor 2 (CCR2),pharmaceutical compositions, and methods for use thereof. Moreparticularly, the CCR2 antagonists are compounds useful for preventing,treating or ameliorating a CCR2 mediated syndrome, disorder or disease.The present invention is further directed to a crystalline succinatesalt of((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone,pharmaceutical compositions containing said salt and the use of saidsalt in the treatment of disorders, such as type II diabetes, obesityand asthma. The present invention is further directed to a novel processfor the preparation of said crystalline succinate salt.

BACKGROUND OF THE INVENTION

CCR2 is a member of the GPCR family of receptors, as are all knownchemokine receptors, and are expressed by monocytes and memoryT-lymphocytes. The CCR2 signaling cascade involves activation ofphospholipases (PLCβ2), protein kinases (PKC), and lipid kinases (PI-3kinase).

Chemoattractant cytokines (i.e., chemokines) are relatively smallproteins (8-10 kD), which stimulate the migration of cells. Thechemokine family is divided into four subfamilies based on the number ofamino acid residues between the first and second highly conservedcysteines.

Monocyte chemotactic protein-1 (MCP-1) is a member of the CC chemokinesubfamily (wherein CC represents the subfamily having adjacent first andsecond cysteines) and binds to the cell-surface chemokine receptor 2(CCR2). MCP-1 is a potent chemotactic factor, which, after binding toCCR2, mediates monocyte and lymphocyte migration (i.e., chemotaxis)toward a site of inflammation. MCP-1 is also expressed by cardiac musclecells, blood vessel endothelial cells, fibroblasts, chondrocytes, smoothmuscle cells, mesangial cells, alveolar cells, T-lymphocytes,marcophages, and the like.

After monocytes enter the inflammatory tissue and differentiate intomacrophages, monocyte differentiation provides a secondary source ofseveral proinflammatory modulators, including tumor necrosis factor-α(TNF-α), interleukin-1 (IL-1), IL-8 (a member of the CXC chemokinesubfamily, wherein CXC represents one amino acid residue between thefirst and second cysteines), IL-12, arachidonic acid metabolites (e.g.,PGE₂ and LTB₄), oxygen-derived free radicals, matrix metalloproteinases,and complement components.

Animal model studies of chronic inflammatory diseases have demonstratedthat inhibition of binding between MCP-1 and CCR2 by an antagonistsuppresses the inflammatory response. The interaction between MCP-1 andCCR2 has been implicated (see Rollins B J, Monocyte chemoattractantprotein 1: a potential regulator of monocyte recruitment in inflammatorydisease, Mol. Med. Today, 1996, 2:198; and Dawson J, et al., Targetingmonocyte chemoattractant protein-1 signaling in disease, Expert Opin.Ther. Targets, 2003 Feb. 7 (1):35-48) in inflammatory diseasepathologies such as psoriasis, uveitis, atherosclerosis, rheumatoidarthritis (RA), multiple sclerosis, Crohn's Disease, nephritis, organallograft rejection, fibroid lung, renal insufficiency, type II diabetesand diabetic complications, diabetic nephropathy, diabetic retinopathy,diabetic retinitis, diabetic microangiopathy, tuberculosis, sarcoidosis,invasive staphylococcia, inflammation after cataract surgery, allergicrhinitis, allergic conjunctivitis, chronic urticaria, ChronicObstructive Pulmonary Disease (COPD), allergic asthma, periodontaldiseases, periodonitis, gingivitis, gum disease, diastoliccardiomyopathies, cardiac infarction, myocarditis, chronic heartfailure, angiostenosis, restenosis, reperfusion disorders,glomerulonephritis, solid tumors and cancers, chronic lymphocyticleukemia, chronic myelocytic leukemia, multiple myeloma, malignantmyeloma, Hodgkin's disease, and carcinomas of the bladder, breast,cervix, colon, lung, prostate, and stomach.

Monocyte migration is inhibited by MCP-1 antagonists (either antibodiesor soluble, inactive fragments of MCP-1), which have been shown toinhibit the development of arthritis, asthma, and uveitis. Both MCP-1and CCR2 knockout (KO) mice have demonstrated that monocyte infiltrationinto inflammatory lesions is significantly decreased. In addition, suchKO mice are resistant to the development of experimental allergicencephalomyelitis (EAE, a model of human MS), cockroach allergen-inducedasthma, atherosclerosis, and uveitis. Rheumatoid arthritis and Crohn'sDisease patients have improved during treatment with TNF-α antagonists(e.g., monoclonal antibodies and soluble receptors) at dose levelscorrelated with decreases in MCP-1 expression and the number ofinfiltrating macrophages.

MCP-1 has been implicated in the pathogenesis of seasonal and chronicallergic rhinitis, having been found in the nasal mucosa of mostpatients with dust mite allergies. MCP-1 has also been found to inducehistamine release from basophils in vitro. During allergic conditions,both allergens and histamines have been shown to trigger (i.e. toup-regulate) the expression of MCP-1 and other chemokines in the nasalmucosa of people with allergic rhinitis, suggesting the presence of apositive feedback loop in such patients.

There remains a need for small molecule CCR2 antagonists for preventing,treating or ameliorating a CCR2 mediated inflammatory syndrome, disorderor disease resulting from MCP-1 induced monocyte and lymphocytemigration to a site of inflammation.

All documents cited herein are incorporated by reference.

SUMMARY OF THE INVENTION

The present invention relates to compounds of Formula (I)

wherein:

A is O, or S;

R⁰ is H, or C₍₁₋₄₎alkyl;

wherein said C₍₁₋₄₎alkyl is optionally substituted with OH,C₍₁₋₄₎alkyl-(OCH₂CH₂)_(n)—OCH₃, OCH₃, CO₂H, C(O)NH₂, SO₂NH₂, orCO₂C₍₁₋₄₎alkyl;

n is 1, 2, or 3;

R¹ is cyclohexyl, or tetrahydropyranyl;

wherein said cyclohexyl or tetrahydropyranyl may be optionallysubstituted with one substituent selected from the group consisting ofOCH₃, OH, CH₂CH₃, —CN, NH₂, NH(CH₃), N(CH₃)₂, and OCF₃;

alternatively, R⁰ and R¹ are taken together with their attached nitrogento form a ring selected from the group consisting of

R^(a) is phenyl; wherein the phenyl is optionally substituted withC(O)NH₂, C(O)NHC₍₁₋₄₎alkyl, SO₂NH₂, C(O)N(C₍₁₋₄₎alkyl)₂, OCH₃, CO₂CH₃,or CO₂H;

R^(b) is C₍₁₋₄₎alkyl, or OC₍₁₋₄₎alkyl;

R² is selected from the group consisting of H, C₍₁₋₄₎alkyl, cyclopropyl,cyclohexyl, phenyl, pyridyl, pyrimidyl, pyrazyl, pyrazolyl, imidazolyl,isoxazolyl, thiazolyl, furyl, and thiophenyl;

wherein said phenyl, pyridyl, pyrimidyl, pyrazyl, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, furyl, or thiophenyl is optionallysubstituted with one substituent selected from the group consisting ofNH₂, NHC₍₁₋₃₎alkyl, N(C₍₁₋₃₎alkyl)₂, C₍₁₋₃₎alkyl, —CN, —CH═CH₂, —CONH₂,—CO₂H, —NO₂, —CONHC₍₁₋₄₎alkyl, CON(C₍₁₋₄₎alkyl)₂, C₍₁₋₄₎alkylCONH₂,—NHCOC₍₁₋₄₎alkyl, —CO₂C₍₁₋₄₎alkyl, CF₃, SO₂C₍₁₋₄₎alkyl, —SO₂NH₂,—SO₂NH(C₍₁₋₄₎alkyl), and —SO₂N(C₍₁₋₄₎alkyl)₂;

R³ is H, or CH₃;

alternatively, R³ and R² are taken together with their attached carbonto form

R⁴ is

R⁵ is H, or CH₃;

alternatively, R⁴ and R⁵ are taken together with their attached nitrogento form a ring selected from the group consisting of

R⁶ is CF₃, or OCF₃;

R⁷ is a CF₃ substituted heteroaryl, provided that R⁷ is not

R_(x) is CF₃, F, Cl, CN, or OCH₃;

R_(y) is H, F, Cl, or CF₃;

R_(z) is H, or F;

and pharmaceutically acceptable salts thereof.

The present invention is further directed to a succinate salt of acompound of formula (I-S)

also known as((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone.In an embodiment of the present invention, the succinate salt of thecompound of formula (I-S) is crystalline. In another embodiment, thepresent invention is directed to a succinate salt of the compound offormula (I-S), wherein the salt is crystalline hydrate form; preferably,the hydrate contains about 0.6 moles water per mole of the compound offormula (I-S). In yet another embodiment of the present invention, thesuccinate salt of the compound of formula (I-S) is crystalline hydrateform containing about 0.6 moles water per mole of the compound offormula (I-S) and is further hygroscopic.

The present invention is further directed to a process for thepreparation of a succinate salt of the compound of formula (I-S),preferably a crystalline succinate salt of the compound of formula(I-S), as described in more detail hereinafter.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates a representative pXRD spectrum for the crystallinesuccinate salt of the compound of formula (I-S).

FIG. 2 illustrates a representative DSC scan for the crystallinesuccinate salt of the compound of formula (I-S)

FIG. 3 illustrates a representative TGA scan for the crystallinesuccinate salt of the compound of formula (I-S)

FIG. 4 illustrates a representative moisture isotherm for thecrystalline succinate salt of the compound of formula (I-S).

FIG. 5 illustrates a DSC thermogram showing conversion of arepresentative sample of the amorphous succinate salt of the compound offormula (I-S) to a crystalline succinate salt of the compound of formula(I-S); and a TGA thermogram for a representative sample of amorphoussuccinate salt of the compound of formula (I-S).

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the compounds of Formula (I)

wherein:

A, R⁰, R¹, R², R³, R⁴ and R⁵ are as defined above.

In an embodiment, the invention is directed to compounds of formula (I)wherein A is O.

In another embodiment, the invention is directed to compounds of formula(I) wherein

A is O, or S;

R⁰ is H, or C₍₁₋₄₎alkyl;

wherein said C₍₁₋₄₎alkyl is optionally substituted with OH,C₍₁₋₄₎alkyl-(OCH₂CH₂)_(n)—OCH₃, or OCH₃;

n is 1, 2, or 3;

R¹ is cyclohexyl, 1-methoxy cyclohex-2-yl, tetrahydropyran-4-yl, or3-methoxy tetrahydropyran-4-yl;

alternatively, R⁰ and R¹ are taken together with their attached nitrogento form a ring selected from the group consisting of

R^(a) is phenyl;

wherein the phenyl is optionally substituted with C(O)NH₂, C(O)NHCH₃,SO₂NH₂, C(O)N(CH₃)₂, OCH₃, CO₂CH₃, or CO₂H;

R² is selected from the group consisting of H, C₍₁₋₄₎alkyl, cyclopropyl,cyclohexyl, phenyl, pyridyl, pyrimidyl, pyrazyl, pyrazolyl, imidazolyl,isoxazolyl, thiazolyl, furyl, and thiophenyl;

wherein said phenyl, pyridyl, pyrimidyl, pyrazyl, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, furyl, or thiophenyl is optionallysubstituted with one substituent selected from the group consisting ofNH₂, NHC₍₁₋₃₎alkyl, N(C₍₁₋₃₎alkyl)₂, C₍₁₋₃₎alkyl, —CN, —CH═CH₂, —CONH₂,—CO₂H, —NO₂, —CONHC₍₁₋₄₎alkyl, CON(C₍₁₋₄₎alkyl)₂, C₍₁₋₄₎alkylCONH₂,—NHCOC₍₁₋₄₎alkyl, —CO₂C₍₁₋₄₎alkyl, CF₃, SO₂C₍₁₋₄₎alkyl, —SO₂NH₂,—SO₂NH(C₍₁₋₄₎alkyl), and —SO₂N(C₍₁₋₄₎alkyl)₂;

R³ is H, or CH₃;

alternatively, R³ and R² are taken together with their attached carbonto form

R⁴ and R⁵ are taken together with their attached nitrogen to form a ringselected from the group consisting of

R⁶ is CF₃, or OCF₃;

R⁷ is

and pharmaceutically acceptable salts thereof.

In another embodiment of the invention

A is O, or S;

R⁰ is H, CH₃, CH₂CH₂CH₂OH, CH₂CH₂OH, CH₂CH₂CH₂(OCH₂CH₂)₃OCH₃, orCH₂CH₂OCH₃;

R¹ is tetrahydropyran-4-yl, or 3-methoxy tetrahydropyran-4-yl;

alternatively, R⁰ and R¹ are taken together with their attached nitrogento form a ring selected from the group consisting of

R^(a) is phenyl;

wherein the phenyl is optionally substituted with C(O)N(CH₃)₂, OCH₃, orCO₂H;

R² is H, C₍₁₋₄₎alkyl, cyclopropyl, cyclohexyl, thiazol-2-yl,1-methyl-imidazol-2-yl, 1-methyl-pyrazol-5-yl, or phenyl;

R³ is H, or CH₃;

alternatively, R³ and R² are taken together with their attached carbonto form

R⁴ and R⁵ are taken together with their attached nitrogen to form a ringselected from the group consisting of

R⁶ is CF₃, or OCF₃;

R⁷ is

and pharmaceutically acceptable salts thereof.

In another embodiment, the present invention is directed to compounds offormula (I) wherein

A is O;

R⁰ is H, CH₃, CH₂CH₂CH₂OH, CH₂CH₂OH, CH₂CH₂CH₂(OCH₂CH₂)₃OCH₃, orCH₂CH₂OCH₃;

R¹ is tetrahydropyran-4-yl, or 3-methoxy tetrahydropyran-4-yl;

alternatively, R⁰ and R¹ may be taken together with their attachednitrogen to form a ring selected from the group consisting of

R^(a) is phenyl;

wherein the phenyl is optionally substituted with C(O)N(CH₃)₂, OCH₃, orCO₂H;

R² is H, C₍₁₋₄₎alkyl, cyclopropyl, cyclohexyl, thiazol-2-yl,1-methyl-imidazol-2-yl, 1-methyl-pyrazol-5-yl, or phenyl;

R³ is H, or CH₃;

alternatively, R³ and R² are taken together with their attached carbonto form

R⁴ and R⁵ are taken together with their attached nitrogen to form a ringselected from the group consisting of

R⁶ is CF₃, or OCF₃;

R⁷ is

and pharmaceutically acceptable salts thereof.

In another embodiment, the present invention is directed to any one ormore compounds, independently selected from the group consisting of:

and pharmaceutically acceptable salts thereof.

In another embodiment, the present invention is directed to a compoundof the formula

or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention relates to a pharmaceuticalcomposition, comprising a compound of formula (I) and a pharmaceuticallyacceptable carrier.

In another embodiment, the invention relates to a pharmaceuticalcomposition made by mixing a compound of formula (I) and apharmaceutically acceptable carrier.

In another embodiment, the invention relates to a process for making apharmaceutical composition comprising mixing a compound of formula (I)and a pharmaceutically acceptable carrier.

The present invention is further directed to a product preparedaccording to any of the processes described herein. In anotherembodiment, the invention relates to the product prepared according tothe process as described in Example 31, which follows herein.

In another embodiment, the present invention is directed to a processfor the preparation of a compound of formula (I), as described in moredetail in the Schemes and Examples which follow herein. In anotherembodiment, the present invention is directed to a process for thepreparation of a compound of formula (I) as described in more detail inExample 31, which follows herein.

In another embodiment, the present invention relates to a compoundselected from the group consisting of

these compounds are useful as intermediates for the preparation ofcompounds of formula (I).

In another embodiment, the invention relates to intermediates useful forthe preparation of a compound of formula (I), more particularly,compounds of formula (XIX)

wherein X is Br, PhSe, or I.

In another embodiment, the present invention relates to a compound offormula (XXI)

useful as an intermediate for the preparation of compounds of formula(I).

In another embodiment, the present invention relates to the process formaking (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and R-(+)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 60% enantiomeric excess.

In another embodiment, the present invention relates to the process formaking (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and R-(+)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 90% enantiomeric excess.

In another embodiment, the present invention relates to the processesfor making (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, as describedabove, wherein the borane reducing complex is selected fromborane-dimethylsulfide complex or borane-N,N-diethylaniline complex.

In another embodiment, the present invention relates to the processesfor making (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, as describedabove, wherein a THF solution of said borane-reducing complex andR-(+)-2-methyl-CBS-oxazaborolidine is added to a solution of the4-dimethoxydihydro-2H-pyran-3(4H)-one in THF.

In another embodiment, the present invention relates to any of theprocesses for making (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, wherein the reaction is carried out in an inertenvironment; in another embodiment of the invention, the inertenvironment is nitrogen gas.

In another embodiment, the present invention relates to any of theprocesses for making (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, further comprising reacting the(S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol with dimethyl sulfate toprovide (R)-3,4,4-trimethoxytetrahydro-2H-pyran.

In another embodiment, the present invention relates to any of theprocesses for making (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, further comprising reacting the(R)-3,4,4-trimethoxytetrahydro-2H-pyran with acid to provide(R)-3-methoxydihydro-2H-pyran-4(3H)-one. In another embodiment, the acidis concentrated hydrochloric acid.

In another embodiment, the present invention relates to the process formaking (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and R-(+)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 60% enantiomeric excess, wherein the reaction is run at atemperature range from 20° C. to 60° C.

In another embodiment, the present invention relates to the process formaking (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and S-(−)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 60% enantiomeric excess.

In another embodiment, the present invention relates to the process formaking (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and S-(−)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol,wherein (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol is formed in at least90% enantiomeric excess.

In another embodiment, the present invention relates to the processesfor making (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, as describedabove, wherein the borane reducing complex is selected fromborane-dimethylsulfide complex or borane-N,N-diethylaniline complex.

In another embodiment, the present invention relates to the processesfor making (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, as describedabove, wherein the THF solution of said borane-reducing complex andS-(−)-2-methyl-CBS-oxazaborolidine is added to a solution of the4-dimethoxydihydro-2H-pyran-3(4H)-one in THF.

In another embodiment, the present invention relates to any of theprocesses for making (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, wherein the reaction is carried out in an inertenvironment; in another embodiment of the invention, the inertenvironment is nitrogen gas.

In another embodiment, the present invention relates to any of theprocesses for making (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, further comprising reacting the(R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol with dimethyl sulfate toprovide (S)-3,4,4-trimethoxytetrahydro-2H-pyran.

In another embodiment, the present invention relates to any of theprocesses for making (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, further comprising reacting the(S)-3,4,4-trimethoxytetrahydro-2H-pyran with acid to provide(S)-3-methoxydihydro-2H-pyran-4(3H)-one. In another embodiment, the acidis concentrated hydrochloric acid.

In another embodiment, the present invention relates to any of theprocesses for making (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol, asdescribed above, comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and S-(−)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 60% enantiomeric excess, wherein the reaction is run at atemperature range from 20° C. to 60° C.

In another embodiment, the invention relates to a method for preventing,treating or ameliorating a CCR2 mediated syndrome, disorder or diseasecomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of formula (I).

In another embodiment, the invention relates to a method for preventing,treating or ameliorating a CCR2 mediated inflammatory syndrome, disorderor disease wherein the syndrome, disorder or disease is associated withelevated MCP-1 expression or MCP-1 overexpression, or is an inflammatorycondition that accompanies syndromes, disorders or diseases associatedwith elevated MCP-1 expression or MCP-1 overexpression comprisingadministering to a subject in need thereof an effective amount of acompound of claim 1.

In another embodiment, the invention relates to a method of preventing,treating or ameliorating a syndrome, disorder or disease, wherein saidsyndrome, disorder or disease is selected from the group consisting of:Chronic Obstructive Pulmonary Disease (COPD), ophthalmic disorders,uveitis, atherosclerosis, rheumatoid arthritis, psoriasis, psoriaticarthritis, atopic dermatitis, multiple sclerosis, Crohn's Disease,ulcerative colitis, nephritis, organ allograft rejection, fibroid lung,renal insufficiency, type-I diabetes, type II diabetes, diabeticcomplications, diabetic nephropathy, diabetic retinopathy, diabeticretinitis, diabetic microangiopathy, overweight, obesity,obesity-associated insulin resistance, metabolic syndrome, tuberculosis,sarcoidosis, invasive staphyloccocia, inflammation after cataractsurgery, allergic rhinitis, allergic conjunctivitis, chronic urticaria,asthma, allergic asthma, periodontal diseases, periodonitis, gingivitis,gum disease, diastolic cardiomyopathies, cardiac infarction,myocarditis, chronic heart failure, angiostenosis, restenosis,reperfusion disorders, aortic abdominal aneurism, glomerulonephritis,solid tumors and cancers, chronic lymphocytic leukemia, chronicmyelocytic leukemia, multiple myeloma, malignant myeloma, Hodgkin'sdisease, and carcinomas of the bladder, breast, cervix, colon, lung,prostate, or stomach and chronic neuroinflammatory disorders including,but not limited to, Alzheimer's disease, ischemic stroke, spinal cordinjury, nerve crush injury and traumatic brain injury comprisingadministering to a subject in need thereof an effective amount of acompound of formula (I).

In another embodiment, the invention relates to a method of preventing,treating or ameliorating a syndrome, disorder or disease, wherein saidsyndrome, disorder or disease is selected from the group consisting of:type I diabetes, type II diabetes, diabetic complications, diabeticnephropathy, diabetic retinopathy, diabetic retinitis, diabeticmicroangiopathy, obesity, obesity-associated insulin resistance,metabolic syndrome, asthma, and allergic asthma, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of formula (I).

In another embodiment, the invention relates to a method of treating adisorder selected from the group consisting of type II diabetes, obesityand asthma comprising administering to a subject in need thereof atherapeutically effective amount of a compound of formula (I).

The present invention is further directed to a succinate salt of acompound of formula (I-S)

wherein the compound of formula (I-S) is also known as((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone.In an embodiment, the succinate salt of the compound of formula (I-S) iscrystalline. In another embodiment, the succinate salt of the compoundof formula (I-S) is a crystalline hydrate form; wherein the hydratecontains about 0.6 moles water per mole of the compound of formula(I-S). In another embodiment of the present invention, the succinatesalt of the compound of formula (I-S) is crystalline hydrate formcontaining about 0.6 moles water per mole of the compound of formula(I-S); wherein the crystalline hydrate form is further hygroscopic.

The present invention is also directed to a crystalline succinate saltof the compound of formula (I-S), wherein the acidic counter-ion issuccinic acid. Additional salt screening was performed on the compoundof formula (I-S), using the following additional acidic counter-ions:HCl acid, sulfuric acid, citric acid, malonic acid, maleic acid,L-tartaric acid, p-toluenesulfonic acid, phosphoric acid and aceticacid. X-ray analysis of the resulting solid residues indicatedcrystalline structures for the sulfate, maleate and phosphate salts; andamorphous structures of the HCl, citrate, malonate, tartrate andtosylate salts.

The crystalline phosphate, sulfate and maleate salts of the compound offormula (I-S) were additional tested in DSC, TGA and moisturesorption/desorption. The sulfate salt showed inter-conversion betweensalt forms and hygroscopic weight increase of 1.6% up to 60% RH and atotal of 26.5% up to 90% RH, with strong hysteresis. The maleate saltshowed a form change and hygroscopic weight increase of 18.3% up to 70%RH and a total of 79.9% up to 90% RH. The phosphate salt showedhygroscopic weight increase of 3.3% up to 60% RH and a total of 69.4% upto 90% RH, with strong hysteresis and deliquescence.

DEFINITIONS

The term “alkyl” refers to both linear and branched chain radicals of upto 12 carbon atoms, preferably up to 6 carbon atoms, unless otherwiseindicated, and includes, but is not limited to, methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl,hexyl, isohexyl, heptyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl,undecyl and dodecyl.

The term “C_((a-b))” (where a and b are integers referring to adesignated number of carbon atoms) refers to an alkyl, alkenyl, alkynyl,alkoxy or cycloalkyl radical or to the alkyl portion of a radical inwhich alkyl appears as the prefix root containing from a to b carbonatoms inclusive. For example, C₍₁₋₄₎ denotes a radical containing 1, 2,3 or 4 carbon atoms.

The term “cycloalkyl” refers to a saturated or partially unsaturatedmonocyclic or bicyclic hydrocarbon ring radical derived by the removalof one hydrogen atom from a single ring carbon atom. Examples ofcycloalkyl radicals include, but are not limited to cyclopropyl,cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,cycloheptyl and cyclooctyl. Additional examples includeC₍₃₋₈₎cycloalkyl, C₍₅₋₈₎cycloalkyl, C₍₃₋₁₂₎cycloalkyl,C₍₃₋₂₀₎cycloalkyl, decahydronaphthalenyl, and2,3,4,5,6,7-hexahydro-1H-indenyl.

The term “boron reducing agent” refers to a boron hydride, often withstabilizers such as ethers, amines, or sulfides. Examples of boronreducing agents include, but are not limited to, borane-tetrahydrofurancomplex, catecholborane, borane-dimethyl aniline complex, andborane-dimethyl sulfide complex.

The term “heteroaryl” refers to a radical derived by the removal of onehydrogen atom from a ring carbon atom of a heteroaromatic ring system. Aheteroaromatic ring system shall denote any five or six memberedmonocyclic aromatic ring structure containing at least one heteroatomselected from the group consisting of O, N and S, optionally containingone to three additional heteroatoms independently selected from thegroup consisting of O, N and S; or a nine or ten membered bicyclicaromatic ring structure containing at least one heteroatom selected fromthe group consisting of O, N and S, optionally containing one to fouradditional heteroatoms independently selected from the group consistingof O, N and S. The heteroaryl group may be attached at any heteroatom orcarbon atom of the ring such that the result is a stable structure.Examples of heteroaryl radicals include, but are not limited to, furyl,thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl,isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl,pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, indolyl,isoindolyl, benzo[b]furyl, benzo[b]thienyl, indazolyl, benzimidazolyl,benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl,cinnolinyl, phthalzinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl,and pteridinyl.

The term “ee” or “enantiomeric excess” is the absolute value of thedifference in mole fractions of a mixture of enantiomers. The molefraction of the (+)- and (−)-enantiomers are expressed as F(+) and F(−)(where F(+)+F(−)=1). The enantiomeric excess is defined as |F(+)−F(−)|.The percent enantiomeric excess is the ee*100. For example a 50/50mixture of (+) and (−) enantiomers has 0% ee, a 5/95 mixture of (+) and(−) enantiomers has a 90% ee, and a 70/30 mixture of (+) and (−)enantiomers has a 40% ee.

The term “inert environment” is a local environment for chemicalreactions which is substantially depleted of atmospheric oxygen andwater vapor. For example, a reaction run under an inert environmentincludes, but is not limited to, reactions run under argon or nitrogenatmosphere.

The term “isolated form” shall mean that the compound is present in aform which is separate from any solid mixture with another compound(s),solvent system or biological environment. In an embodiment, the presentinvention is directed to a succinate salt of the compound of formula(I-S), preferably a crystalline succinate salt of the compound offormula (I-S), wherein the salt is present and/or prepared as anisolated form.

The term “substantially free of other salt form(s)” when used todescribe the succinate salt of the compound of formula (I-S) shall meanthat mole percent of any other salt form(s) in the isolated succinatesalt of the compound of formula (I-S) is less than about 5 mole percent,preferably less than about 2 mole percent, more preferably, less thanabout 0.5 mole percent, most preferably less than about 0.1 molepercent. In an embodiment, the present invention is directed to asuccinate salt of the compound of formula (I-S), preferably acrystalline succinate salt of the compound of formula (I-S), wherein thesalt is present and/or prepared as form which is substantially free ofother salt form(s).

The term “substantially pure form” shall mean that the mole percent ofimpurities in the isolated compound is less than about 5 mole percent,preferably less than about 2 mole percent, more preferably, less thanabout 0.5 mole percent, most preferably, less than about 0.1 molepercent. In an embodiment, the present invention is directed to asuccinate salt of the compound of formula (I-S), preferably acrystalline succinate salt of the compound of formula (I-S), wherein thesalt is present and/or prepared as a substantially pure form.

For use in medicines, the salts of the compounds of this invention referto non-toxic “pharmaceutically acceptable salts.” FDA approvedpharmaceutically acceptable salt forms (Ref International J. Pharm.1986, 33, 201-217; J. Pharm. Sci., 1977, January, 66(1), p1) includepharmaceutically acceptable acidic/anionic or basic/cationic salts.

Throughout this specification, compounds are described as beingseparated, usually by silica gel column, although preporatory thin layerchromatography, or high or low pressure liquid choromatography may alsobe used. It is generally accepted that when eluting compounds through asilica gel-type separation medium, that the least polar compounds elutebefore the more polar compounds. Therefore, the term “less polarisomer”, refers to the isomer that will elute first from a silica geltype separation medium.

Abbreviations

Herein and throughout this application, the following abbreviations maybe used.

-   -   AIBN azobisisobutyronitrile    -   BOC or Boc tert-butyloxycarbonyl    -   DCC dicyclohexylcarbodiimide    -   DCM dicholomethane    -   EDCI or EDC 1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide    -   DIAD diisopropylazodicarboxylate    -   DIEA diisopropylethylamine    -   DSC differential scanning calorimetry    -   Et ethyl    -   EtOAc ethyl acetate    -   ee enantiomeric excess    -   eq equivalents    -   HOBt hydroxybenzotriazole    -   LiHMDS lithium bis(trimethylsilyl)amide    -   M moles/liter    -   Me methyl    -   MIBK methyl isobutyl ketone    -   min. minutes    -   n-BuLi n-butyl lithium    -   NBS or NIS N-bromo succinimide or N-iodo succinimide    -   OAc acetate    -   Ph phenyl    -   PyBrop bromo-tris-pyrrolidinophosphonium hexafluorophosphate    -   RH relative humidity    -   rt room temperature    -   TBAF tetrabutylammonium fluoride    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran    -   TBS or TBDMS tertbutyldimethylsilyl    -   TLC thin layer chromatography

Pharmaceutically acceptable acidic/anionic salts include, and are notlimited to acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate,bromide, calcium edetate, camsylate, carbonate, chloride, citrate,dihydrochloride, edetate, edisylate, estolate, esylate, fumarate,glyceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate,hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide,isethionate, lactate, lactobionate, malate, maleate, mandelate,mesylate, methylbromide, methylnitrate, methylsulfate, mucate,napsylate, nitrate, pamoate, pantothenate, phosphate/diphosphate,polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate,tannate, tartrate, teoclate, tosylate and triethiodide. Organic orinorganic acids also include, and are not limited to, hydriodic,perchloric, sulfuric, phosphoric, propionic, glycolic, methanesulfonic,hydroxyethanesulfonic, oxalic, 2-naphthalenesulfonic, p-toluenesulfonic,cyclohexanesulfamic, saccharinic or trifluoroacetic acid.

Pharmaceutically acceptable basic/cationic salts include, and are notlimited to aluminum, 2-amino-2-hydroxymethyl-propane-1,3-diol (alsoknown as tris(hydroxymethyl)aminomethane, tromethane or “TRIS”),ammonia, benzathine, t-butylamine, calcium, calcium gluconate, calciumhydroxide, chloroprocaine, choline, choline bicarbonate, cholinechloride, cyclohexylamine, diethanolamine, ethylenediamine, lithium,LiOMe, L-lysine, magnesium, meglumine, NH₃, NH₄OH, N-methyl-D-glucamine,piperidine, potassium, potassium-t-butoxide, potassium hydroxide(aqueous), procaine, quinine, sodium, sodium carbonate,sodium-2-ethylhexanoate (SEH), sodium hydroxide, triethanolamine orzinc.

Methods of Use

The present invention is directed to a method for preventing, treatingor ameliorating a CCR2 mediated syndrome, disorder or disease comprisingadministering to a subject in need thereof an effective amount of acompound of Formula (I) or a form, composition or medicament thereof.

Examples of a CCR2 mediated syndrome, disorder or disease for which thecompounds of Formula (I) are useful include chronic obstructivepulmonary disorder (COPD), ophthalmic disorders, uveitis,atherosclerosis, rheumatoid arthritis, psoriasis, psoriatic arthritis,atopic dermatitis, multiple sclerosis, Crohn's Disease, ulcerativecolitis, nephritis, organ allograft rejection, fibroid lung, renalinsufficiency, type-I diabetes, type II diabetes, diabeticcomplications, diabetic nephropathy, diabetic retinopathy, diabeticretinitis, diabetic microangiopathy, overweight, obesity,obesity-associated insulin resistance, metabolic syndrome, tuberculosis,chronic obstructive pulmonary disease, sarcoidosis, invasivestaphyloccocia, inflammation after cataract surgery, allergic rhinitis,allergic conjunctivitis, chronic urticaria, asthma, allergic asthma,periodontal diseases, periodonitis, gingivitis, gum disease, diastoliccardiomyopathies, cardiac infarction, myocarditis, chronic heartfailure, angiostenosis, restenosis, reperfusion disorders, aorticabdominal aneurism, multiple sclerosis, glomerulonephritis, solid tumorsand cancers, chronic lymphocytic leukemia, chronic myelocytic leukemia,multiple myeloma, malignant myeloma, Hodgkin's disease, carcinomas ofthe bladder, breast, cervix, colon, lung, prostate, or stomach, andchronic neuroinflammatory disorders including, but not limited to,Alzheimer's disease, ischemic stroke, spinal cord injury, nerve crushinjury and traumatic brain injury.

Some of the quantitative expressions given herein are qualified with theterm “about”. It is understood that whether the term “about” is usedexplicitly or not, every quantity given herein is meant to refer to boththe actual given value and the approximation to such given value thatwould reasonably be inferred based on the ordinary skill in the art,including approximations due to the experimental and/or measurementconditions for such given value. In addition, some of the quantitativeexpressions herein are recited as a range from about amount X to aboutamount Y. It is understood that wherein a range is recited, the range isnot limited to the recited upper and lower bounds, but rather includesthe full range from about amount X through about amount Y, or any rangetherein.

The term “administering” with respect to the methods of the invention,means a method for therapeutically or prophylactically preventing,treating or ameliorating a syndrome, disorder or disease as describedherein by using a compound of Formula (I) or a form, composition ormedicament thereof. Such methods include administering an effectiveamount of said compound, compound form, composition or medicament atdifferent times during the course of a therapy or concurrently in acombination form. The methods of the invention are to be understood asembracing all known therapeutic treatment regimens.

The term “subject” refers to a patient, which may be animal, typically amammal, typically a human, which has been the object of treatment,observation or experiment. In one aspect of the invention, the subjectis at risk of (or susceptible to) developing a syndrome, disorder ordisease that is associated with elevated MCP-1 expression or MCP-1overexpression, or a patient with an inflammatory condition thataccompanies syndromes, disorders or diseases associated with elevatedMCP-1 expression or MCP-1 overexpression.

The term “therapeutically effective amount” means that amount of activecompound or pharmaceutical agent that elicits the biological ormedicinal response in a tissue system, animal or human, that is beingsought by a researcher, veterinarian, medical doctor, or otherclinician, which includes preventing, treating or ameliorating thesymptoms of a syndrome, disorder or disease being treated.

The term “uveitis” generically refers to any inflammatory diseaseinvolving the eye. Uveitis can be divided into clinically distinctsubtypes based on the part of the eye in which the inflammation ispresent (percentages correspond to patients known to fit thesecategories): anterior (51%), intermediate (13%), posterior (20%), orpanuveitis (16%) and, according to the course of the disease, as eitheracute (16%), recurring (26%), or chronic (58%). Those with anterioruveitis (0.19%) eventually develop irreparable vision damage despiteaggressive treatment such as unilateral blindness (9%), bilateralblindness (2%), or unilateral or bilateral vision impairment (8%). Mostcases of uveitis are idiopathic, but known causes include infection(e.g., toxoplasmosis, cytomegalovirus, and the like) or development as acomponent of a systemic inflammatory and/or autoimmune disorder (e.g.,juvenile RA, HLA-B27 associated spondyloarthropathies, sarcoidosis, andthe like). (HLA-B27: Human Leukocyte Antigen B*27—is a class I surfaceantigen encoded by the B locus in the major histocompatibility complex(MHC) on chromosome 6 and presents microbial antigens to T cells.HLA-B27 is strongly associated with a certain set of autoimmune diseasesreferred to as the seronegative spondyloarthropathies.)

When employed as CCR2 inhibitors, the compounds of the invention may beadministered in an effective amount within the dosage range of about 0.5mg to about 10 g, or any amount or range therein, preferably betweenabout 0.5 mg to about 5 g, or any amount or range therein, in single ordivided daily doses. The dosage administered will be affected by factorssuch as the route of administration, the health, weight and age of therecipient, the frequency of the treatment and the presence of concurrentand unrelated treatments.

It is also apparent to one skilled in the art that the therapeuticallyeffective dose for compounds of the present invention or apharmaceutical composition thereof will vary according to the desiredeffect. Therefore, optimal dosages to be administered may be readilydetermined by one skilled in the art and will vary with the particularcompound used, the mode of administration, the strength of thepreparation, and the advancement of the disease condition. In addition,factors associated with the particular subject being treated, includingsubject age, weight, diet and time of administration, will result in theneed to adjust the dose to an appropriate therapeutic level. The abovedosages are thus exemplary of the average case. There can, of course, beindividual instances where higher or lower dosage ranges are merited,and such are within the scope of this invention.

The compounds of Formula (I) may be formulated into pharmaceuticalcompositions comprising any known pharmaceutically acceptable carriers.Exemplary carriers include, but are not limited to, any suitablesolvents, dispersion media, coatings, antibacterial and antifungalagents and isotonic agents. Exemplary excipients that may also becomponents of the formulation include fillers, binders, disintegratingagents and lubricants.

The pharmaceutically-acceptable salts of the compounds of Formula (I)include the conventional non-toxic salts or the quaternary ammoniumsalts which are formed from inorganic or organic acids or bases.Examples of such acid addition salts include acetate, adipate, benzoate,benzenesulfonate, citrate, camphorate, dodecylsulfate, hydrochloride,hydrobromide, lactate, maleate, methanesulfonate, nitrate, oxalate,pivalate, propionate, succinate, sulfate and tartrate. Base saltsinclude ammonium salts, alkali metal salts such as sodium and potassiumsalts, alkaline earth metal salts such as calcium and magnesium salts,salts with organic bases such as dicyclohexylamino salts and salts withamino acids such as arginine. Also, the basic nitrogen-containing groupsmay be quaternized with, for example, alkyl halides.

The pharmaceutical compositions of the invention may be administered byany means that accomplish their intended purpose. Examples includeadministration by parenteral, subcutaneous, intravenous, intramuscular,intraperitoneal, transdermal, buccal or ocular routes. Alternatively orconcurrently, administration may be by the oral route. Suitableformulations for parenteral administration include aqueous solutions ofthe active compounds in water-soluble form, for example, water-solublesalts, acidic solutions, alkaline solutions, dextrose-water solutions,isotonic carbohydrate solutions and cyclodextrin inclusion complexes.

The present invention also encompasses a method of making apharmaceutical composition comprising mixing a pharmaceuticallyacceptable carrier with any of the compounds of the present invention.Additionally, the present invention includes pharmaceutical compositionsmade by mixing a pharmaceutically acceptable carrier with any of thecompounds of the present invention. As used herein, the term“composition” is intended to encompass a product comprising thespecified ingredients in the specified amounts, as well as any productwhich results, directly or indirectly, from combinations of thespecified ingredients in the specified amounts.

Polymorphs and Solvates

Furthermore, the compounds of the present invention may have one or morepolymorph or amorphous crystalline forms and as such are intended to beincluded in the scope of the invention. In addition, the compounds mayform solvates, for example with water (i.e., hydrates) or common organicsolvents. As used herein, the term “solvate” means a physicalassociation of the compounds of the present invention with one or moresolvent molecules. This physical association involves varying degrees ofionic and covalent bonding, including hydrogen bonding. In certaininstances the solvate will be capable of isolation, for example when oneor more solvent molecules are incorporated in the crystal lattice of thecrystalline solid. The term “solvate” is intended to encompass bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.

It is intended that the present invention include within its scopepolymorphs and solvates of the compounds of the present invention. Thus,in the methods of treatment of the present invention, the term“administering” shall encompass the means for treating, ameliorating orpreventing a syndrome, disorder or disease described herein with thecompounds of the present invention or a polymorph or solvate thereof,which would obviously be included within the scope of the inventionalbeit not specifically disclosed.

In another embodiment, the invention relates to a compound as describedin the Examples of Formula (I) for use as a medicament.

In another embodiment, the invention relates to the use of a compound asdescribed in the Examples of Formula (I) for the preparation of amedicament for the treatment of a disease associated with an elevated orinappropriate CCR2 activity.

The present invention includes within its scope prodrugs of thecompounds of this invention. In general, such prodrugs will befunctional derivatives of the compounds which are readily convertible invivo into the required compound. Thus, in the methods of treatment ofthe present invention, the term “administering” shall encompass thetreatment of the various disorders described with the compoundspecifically disclosed or with a compound which may not be specificallydisclosed, but which converts to the specified compound in vivo afteradministration to the patient. Conventional procedures for the selectionand preparation of suitable prodrug derivatives are described, forexample, in “Design of Prodrugs”, Ed. H. Bundgaard, Elsevier, 1985.

Where the compounds according to this invention have at least one chiralcenter, they may accordingly exist as enantiomers. Where the compoundspossess two or more chiral centers, they may additionally exist asdiastereomers. It is to be understood that all such isomers and mixturesthereof are encompassed within the scope of the present invention.

Where the processes for the preparation of the compounds according tothe invention give rise to mixture of stereoisomers, these isomers maybe separated by conventional techniques such as preparativechromatography. The compounds may be prepared in racemic form, orindividual enantiomers may be prepared either by enantiospecificsynthesis or by resolution. The compounds may, for example, be resolvedinto their component enantiomers by standard techniques, such as theformation of diastereomeric pairs by salt formation with an opticallyactive acid, such as (−)-di-p-toluoyl-D-tartaric acid and/or(+)-di-p-toluoyl-L-tartaric acid followed by fractional crystallizationand regeneration of the free base. The compounds may also be resolved byformation of diastereomeric esters or amides, followed bychromatographic separation and removal of the chiral auxiliary.Alternatively, the compounds may be resolved using a chiral HPLC column.

During any of the processes for preparation of the compounds of thepresent invention, it may be necessary and/or desirable to protectsensitive or reactive groups on any of the molecules concerned. This maybe achieved by means of conventional protecting groups, such as thosedescribed in Protective Groups in Organic Chemistry, ed. J. F. W.McOmie, Plenum Press, 1973; and T. W. Greene & P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, John Wiley & Sons, 1991. The protectinggroups may be removed at a convenient subsequent stage using methodsknown from the art.

Preparation of Crystalline Succinate Salt of the Compound of Formula(I-S)

The crystalline succinate salt of the compound of formula (I-S) of thepresent invention may be prepared from the corresponding amorphoussuccinate salt of the compound of formula (I-S) by heating to atemperature in the range of from about 140° C. to about 150° C.,preferably to about 140° C., and then cooling to about room temperatureto effect crystallization, as described in more detail in Example 52,which follows herein.

Alternatively, the crystalline succinate salt of the compound of formula(I-S) of the present invention may be prepared from the correspondingamorphous succinate salt of the compound of formula (I-S) bycrystallization from a suitably selected solvent such as methyl isobutylketone (MIBK), as described in more detail in Example 53, which followherein. For the crystallization of the succinate salt of the compound offormula (I-S) the suitably selected solvent is other than water,methanol, ethanol, acetone, acetonitrile. isopropyl acetate,nitromethane, tetrahydrofuran, methyl ethyl ketone, dichloromethane,toluene, methyl isopropyl ketone (MIPK).

The amorphous succinate salt of the compound of formula (I-S) may beprepared, for example, as described in Example 30, which follows herein.

Powder X-Ray Diffraction (pXRD)

The succinate salt of the compound of formula (I-S) was characterized asto its powder X-ray diffraction pattern (pXRD), in example as follows.The sample was examined using an X-ray diffractometer (Philips ModelX′PERT PRO PW3040) with X'Celerator detector and graded multilayerparabolic X-ray mirror. The samples were scanned from 3 to 40° 2θ, at astep size 0.0165° 2θ and a time per step of 2000.025 seconds. The tubevoltage and current were 45 KV and 40 mA, respectively. The sample waspacked on a zero background XRD-holder and scanned under ambienttemperature and humidity conditions.

A pXRD spectrum was measured for a representative sample of thecrystalline succinate salt of the compound of formula (I-S), as shown inFIG. 1. In an embodiment, the crystalline succinate salt of the compoundof formula (I-S) may be characterized by its powder X-ray diffractionpattern, which comprised the peaks listed in Table 1, below.

TABLE 1 pXRD Peaks: Crystalline Succinate Salt of Compound of Formula(I-S) Position [°2θ] d-spacing [Å] Relative Intensity [%] 7.27 12.17 310.03 8.82 4 10.51 8.42 4 11.27 7.85 8 12.26 7.22 2 13.87 6.38 67 14.346.18 21 14.63 6.05 16 15.96 5.55 17 16.73 5.30 11 17.66 5.02 13 18.334.84 16 19.22 4.62 100 19.78 4.49 36 20.11 4.42 36 20.86 4.26 26 21.344.16 20 22.01 4.04 50 22.67 3.92 16 23.62 3.77 15 24.14 3.69 8 25.783.46 17 27.07 3.29 14 27.64 3.23 7 28.40 3.14 6 28.97 3.08 4 30.30 2.954 31.91 2.80 3 33.34 2.69 4 35.91 2.50 4

In an embodiment, the crystalline succinate salt of the compound offormula (I-S) is characterized by its pXRD pattern which comprises peakshaving a relative intensity greater than or equal to about 3%, as listedin Table 2, below.

TABLE 2 pXRD Peaks: Crystalline Succinate Salt of Compound of Formula(I-S) Position [°2θ] d-spacing [Å] Relative Intensity [%] 10.03 8.82 410.51 8.42 4 11.27 7.85 8 12.26 7.22 2 13.87 6.38 67 14.34 6.18 21 14.636.05 16 15.96 5.55 17 16.73 5.30 11 17.66 5.02 13 18.33 4.84 16 19.224.62 100 19.78 4.49 36 20.11 4.42 36 20.86 4.26 26 21.34 4.16 20 22.014.04 50 22.67 3.92 16 23.62 3.77 15 24.14 3.69 8 25.78 3.46 17 27.073.29 14 27.64 3.23 7 28.40 3.14 6 28.97 3.08 4 30.30 2.95 4 33.34 2.69 435.91 2.50 4

In an embodiment, the crystalline succinate salt of the compound offormula (I-S) is characterized by its pXRD pattern which comprises peakshaving a relative intensity greater than or equal to about 5%, as listedin Table 3, below.

TABLE 3 pXRD Peaks: Crystalline Succinate Salt of Compound of Formula(I-S) Position [°2θ] d-spacing [Å] Relative Intensity [%] 11.27 7.85 812.26 7.22 2 13.87 6.38 67 14.34 6.18 21 14.63 6.05 16 15.96 5.55 1716.73 5.30 11 17.66 5.02 13 18.33 4.84 16 19.22 4.62 100 19.78 4.49 3620.11 4.42 36 20.86 4.26 26 21.34 4.16 20 22.01 4.04 50 22.67 3.92 1623.62 3.77 15 24.14 3.69 8 25.78 3.46 17 27.07 3.29 14 27.64 3.23 728.40 3.14 6

In an embodiment, the crystalline succinate salt of the compound offormula (I-S) is characterized by its pXRD pattern which comprises peakshaving a relative intensity greater than or equal to about 10%, aslisted in Table 4, below.

TABLE 4 pXRD Peaks: Crystalline Succinate Salt of Compound of Formula(I-S) Position [°2θ] d-spacing [Å] Relative Intensity [%] 13.87 6.38 6714.34 6.18 21 14.63 6.05 16 15.96 5.55 17 16.73 5.30 11 17.66 5.02 1318.33 4.84 16 19.22 4.62 100 19.78 4.49 36 20.11 4.42 36 20.86 4.26 2621.34 4.16 20 22.01 4.04 50 22.67 3.92 16 23.62 3.77 15 25.78 3.46 1727.07 3.29 14

In another embodiment, the present invention is directed to acrystalline succinate salt of the compound of formula (I-S) ascharacterized by the following pXRD peak, listed in ° 2θ: 10.03, 10.51,11.27, 13.87, 19.22 and 22.01. In another embodiment, the presentinvention is directed to a crystalline succinate salt of the compound offormula (I-S) as characterized by the following pXRD peak, listed in °20: 11.27, 13.87, 19.22 and 22.01.

Differential Scanning Calorimetry (DSC)

The crystalline succinate salt of the compound of formula (I-S) wasfurther subjected to DSC analysis. A representative sample was testedusing a TA Instruments Model Q100O differential scanning calorimeter.The sample was analyzed as received in an open aluminum pan. The DSC wasprogrammed to heat from 25° C. to 300° C. at a heating rate of 10°C./min with a nitrogen purge.

Thermal analysis (via DSC scanning) was completed for a representativesample of the crystalline succinate salt of the compound of formula(I-S), as shown in FIG. 2. The crystalline succinate salt of thecompound of formula (I-S) exhibited an onset melting temperature ofabout 156° C., a peak temperature of melting of about 158° C. and anenthalpy 68.3 J/g.

Thermogravimetric Analysis (TGA)

The crystalline succinate salt of the compound of formula (I-S) wasfurther subjected to TGA analysis. A representative sample was tested,as received, for total weight loss using a TA Instruments Model Q5000IRTGA thermogravimetric calorimeter. The sample was placed in a tarredaluminum pan, automatically weighed and inserted into the TGA furnace.The sample was scanned from 25° C. to 300° C. at a heating rate of 10°C./min with a 90 mL/min nitrogen purge and a 10 mL/min helium balancepurge.

A TGA trace was measured for a representative sample of the crystallinesuccinate salt of the compound of formula (I-S), as shown in FIG. 3. A1.8% weight loss was observed between room temperature and 144° C., dueto dehydration/desolvation; followed by decomposition at 151° C. Theseresults indicate that the crystalline succinate salt of the compound offormula (I-S) is hydrate; wherein the hydrate contains about 0.6 molesof water per mole of the compound of formula (IS).

Moisture Isothermal Analysis

The crystalline succinate salt of the compound of formula (I-S) wasfurther subjected to moisture sorption analysis. The moisture sorptionanalysis was performed using Hiden Isochema system Model IGAsorp. Thesample (˜5 mg) was run in a stainless-steel mesh crucible. The samplewas initially dried at 60° C. for 30 minutes, then the moisture profilewas evaluated by monitoring vapor adsorption/desorption over the rangeof 0% RH to 90% RH at 25° C. The moisture profile consisted of 2 cyclesof vapor adsorption/desorption.

FIG. 4 illustrates two cycles of vapor sorption/desorption for arepresentative sample of the crystalline succinate salt of the compoundof formula (I-S). The weight of the sample increased 3.8% up to 70% RH,with a total of 70% uptake up to 90% RH. Further, strong hysteresis wasobserved in the multiple sorption/desorption cycles with about 2.7%moisture (equivalent to 0.9 moles of water) retained at the end of thedesorption phase. Thus, the crystalline succinate salt of the compoundof formula (I-S) is hygroscopic.

Solubility

The crystalline succinate salt of the compound of formula (I-S) wastested for solution solubility and measured to be soluble at >50 mg/mlin water and at >100 mg/ml in 0.1N NaOH, pH 2, pH4 and pH6 citratebuffers; in pH 8 and pH 10 borate buffers; simulated intestinal fluid,simulated gastric fluid; 0.5% methocel and 20% HpbCD.

General Reaction Scheme

Representative compounds of the present invention can be synthesized inaccordance with the general synthetic methods described below. Compoundsof Formula (I) can be prepared by methods known to those who are skilledin the art. The following reaction schemes are only meant to representexamples of the invention and are in no way meant to be a limit of theinvention.

Compounds of Formula (I) wherein A is O and wherein R⁰, R² and R³ areeach hydrogen may be prepared according to the processes outlined inScheme 1.

Scheme 1 illustrates a synthetic route leading to compounds of Formula(I) wherein R⁰, R² and R³ are each hydrogen. Commercially availableesters of formula (II) is alkylated by reacting with a suitable selectedbase such as LiHMDS, and the like; and then reacted witht-butyl(2-iodoethoxy)dimethylsilane, in an organic solvent such as THFor diethyl ether, at a temperature in the range of −78° C. to 20° C. Theresulting alkylated ester is then saponified by reacting with an aqueousbase such as NaOH, KOH, or LiOH, in a solvent such as methanol orethanol, at a temperature in the range of 0° C. to 60° C., to yield thecorresponding acid, a compound of formula (III).

The acid of formula (III) is then reacted with a suitably selected,commercially available amine of formula (IV), in the presence of acoupling reagent such as EDCI/HOBt, PyBrop or DCC, in an organic solventsuch as THF, dichloromethane or 1,2-dichloroethane, at a temperature inthe range of about 0° C. to about 25° C., to yield the correspondingamide; which is then reacted to de-protect the alcohol functionality, byreacting with a fluoride source such as TBAF, in a solvent such as THF,at a temperature range of about 0° C. to 60° C., to yield thecorresponding, desilylated alcohol of formula (V).

The alcohol of formula (V) is reacted with a reagent such asN-(phenylseleno)phthalimide (VI) or phenylselenyl chloride, and a LewisAcid such as borontrifluoride etherate, in a solvent such as DCM or1,2-dichloroethane, at a temperature of 0° C. to 50° C., to yield thecorresponding cyclic ether of formula (VII).

The cyclic ether (VII) is reacted with a suitably selected reducingagent such as tri-n-butylstannane or tris(trimethylsilyl)silane, in thepresence of a radical initiator such as AIBN, in a solvent such asbenzene or toluene, at a temperature of 60° C. to 120° C.; and theresulting intermediate, de-protected at the N-Boc carbamate by reactingwith an acid such as TFA or HCl, in a solvent such as DCM, acetonitrile,THF, or dioxane, at a temperature of 0° C. to 80° C., to yield thecorresponding amine of formula (VIII).

The amine of formula (VIII) is reacted with a suitably substitutedketone of formula (IX), in the presence of a suitably selected reducingreagent such as NaBH₄, NaBH₃CN or NaBH(OAc)₃, in an organic base such astriethylamine, diisopropylethylamine or N-methylmorpholine, with orwithout molecule sieves, in an organic solvent such as dichloromethane,1,2-dichloroethane or THF, at a temperature in the range of 0° C. toabout 25° C., to yield the corresponding compound of formula (I) whereinR⁰, R² and R³ are each hydrogen.

Those skilled in the art will recognize that compounds of formula (I)where R₀ is not H may be synthesized from compounds of formula (I) whereR₀ is H, by standard alkylation procedures. Such alkylation proceduresinclude, but are not limited to, reductive alkylation. For example, acompound of formula (I) where R₀ is H may be dissolved in a solvent,such as THF, and reacted with an aldehyde and a reductant, such assodium triacetoxyborohydride. Suitable temperatures include a rangeextending from about 25° C. to 50° C.

Compounds of Formula (X) may be prepared according to the processesoutlined in Scheme 2.

An alcohol of formula (V), prepared for example, as described in Scheme1 above, is converted to the corresponding aldehyde by reacting with asuitably selected oxidant such as Dess-Martin periodinane, Swern, orpyridinium chlorochromate, in a solvent such as DCM or1,2-dichloroethane, at a temperature of 0° C. to 50° C. The resultingintermediate is then reacted with a suitably selected Grignard reagentor lithium reagent of formula (XI), in a solvent such as diethyl ether,THF, or toluene, at a temperature of −78° C. to 25° C., to yield thecorresponding alcohol of formula (X).

Compounds of Formula (I) wherein A is O and wherein R² is other thanhydrogen may be prepared from the corresponding compound of Formula (X),by substituting said compound of formula (X) for the compound of formula(V) in Scheme 1, above.

Compounds of Formula (XII) may be prepared according to the processesoutlined in Scheme 3.

An alcohol of formula (V), prepared for example as described in Scheme1, above, is reacted with a suitably selected oxidant such as CrO₃,Ru/NaIO₄, or KMnO₄, in a solvent such as acetone or water, at atemperature of 0° C. to 50° C., to yield the corresponding carboxylicacid of formula (XIII).

The acid of formula (XIII) is reacted with a suitably selectedalkylating agent such as trimethylsilyl(diazomethane) in a solvent suchas methanol or ethanol, at a temperature of −20° C. to 25° C., to yieldthe corresponding methyl ester of formula (XIV).

The methyl ester of formula (XIV) is reacted with a lithium or Grignardreagent (XI) in a solvent such as diethyl ether, THF, or toluene, at atemperature of −78° C. to 25° C., to yield the substituted alcohol(XII).

Compounds of Formula (I) wherein A is O and wherein R² and R³ are otherthan hydrogen may be prepared from the corresponding compound of Formula(XII), by substituting said compound of formula (XII) for the compoundof formula (V) in Scheme 1, above.

Compounds of Formula (XV) may be prepared according to the processesoutlined in Scheme 4.

A suitably selected acid of formula (XIII), prepared for example asdescribed in Scheme 3 above, is reacted with a reagent such asN-(phenylseleno)phthalimide (VI) or phenylselenyl chloride, and a LewisAcid such as borontrifluoride etherate, in a solvent such as DCM or1,2-dichloroethane, at a temperature of 0° C. to 50° C., to yield thecorresponding bicyclic lactone of formula (XV).

Compounds of Formula (I) wherein A is O and wherein R² and R³ are takentogether with the carbon atom to which they are bound to form C═O may beprepared from the corresponding compound of Formula (XV) by substitutingsaid compound of formula (XV) for the compound of formula (V) in Scheme1, above.

Compounds of Formula (XVI) may be prepared according to the processesoutlined in Scheme 5.

An alcohol of formula (V), prepared for example as described in Scheme1, above, is reacted with a thioacid such as thioacetic acid, in thepresence of a phosphine such as triphenylphosphine or tributylphosphineand in the presence of an activating agent such asdiisopropylazodicarboxylate (DIAD) or diethylazodicarboxylate (DEAD), ina solvent such as THF, diethyl ether, DCM or 1,2-dichloroethane, at atemperature of 0° C. to 60° C. (i.e. under Mitsunobu conditions), toyield the corresponding thioester of formula (XVII).

The thioester of formula (XVII) is reacted with an aqueous base such asNaOH, LiOH or KOH, in a solvent such as methanol or ethanol, at atemperature of 0° C. to 60° C., to yield the corresponding thiol offormula (XVIII).

The thiol of formula (XVIII) is reacted with a radical initiator such asAIBN, in a solvent such as benzene or toluene, at a temperature of 60°C. to 120° C., to yield the corresponding bicyclic thioether of formula(XVI).

Compounds of formula (I) wherein A is S, R⁰ is hydrogen, and wherein R₂and R₃ are other than hydrogen may be prepared according to theprocedures of Scheme 5, by starting with compound (XII) (as prepared inScheme 3) in place of compound (V). Additionally, reductive alkylationas described in Scheme 1 can be employed by one of ordinary skill in theart as a means for transforming these compounds of formula (I) where Ais S and R₀ is H to compounds of formula (I) where A is S and R₀ isother than H.

Compounds of Formula (VIII) may alternatively be prepared according tothe procedure of Scheme 6.

A compound of formula (III), prepared for example as described in Scheme1 above, is cyclized by reacting with a suitably selected de-silylatingagent, such as tetra-butyl ammonium fluoride, and the like, in a solventsuch as THF, and the like, at a temperature range from about −20° C. toabout 50° C. to yield the corresponding lactone of formula (XVII).

The lactone of formula (XVII) is reacted with a suitably selectedreducing agent, such as NaBH₄, LiAlH₄, and the like, in a suitablyselected solvent, such as THF, and the like, at a temperature range offrom about −20° C. to about 50° C., to yield the corresponding diol offormula (XVIII).

The diol of formula (XVIII) is reacted with a suitably selectedhalogenating reagent such as N-bromo-succinimide, N-iodo-succinimide,Br₂, and the like, in a solvent such as THF, EtOAc, CH₂Cl₂, and thelike, at a temperature range of from about 0° C. to about 100° C., toyield the corresponding intermediate of formula (XIX).

The intermediate of formula (XIX) is hydrogenated by reacted withhydrogen gas, in a solvent or mixture of solvents, such as THF, EtOAc,methanol, and the like, in the presence of a suitably selected catalystsuch as Pd/C, Pt/C, and the like, at about room temperature, to yieldthe corresponding alcohol of formula (XX).

Alternatively, the intermediate of formula (XIX) may be reacted with areducing agent such as tri-n-butylstannane, tris(trimethylsilyl)silane,and the like, in the presence of a radical initiator such as AIBN, andthe like, in a solvent such as benzene, toluene, and the like, at atemperature in the range of from about 60° C. to about 120° C. to yieldthe corresponding alcohol of formula (XX).

The alcohol of formula (XX) is reacted with a suitably selected oxidantsuch as CrO₃, Ru/NaIO₄, KMnO₄ and the like, in a solvent such asacetone, water, and the like, at a temperature in the range of fromabout 0° C. to about 50° C., to yield the corresponding carboxylic acidof formula (XXI).

The carboxylic acid of formula (XXI) is reacted with a suitably selectedamine of formula (IV), in the presence of a coupling reagent such asEDCI/HOBt, PyBrop, DCC, and the like, in an organic solvent such as THF,dichloromethane, 1,2-dichloroethane, and the like, at a temperature inthe range of about 0° C. to about 25° C., to yield the correspondingamide of formula (XXII).

The amide of formula (XXII) is de-protected by reacting under acidicconditions, such as HCl in MeOH, at temperature ranging from about 25°C. to about 80° C., to yield the corresponding compound of formula(VIII), which may then be reacted, as described in Scheme 1 above, toyield the corresponding compound of Formula (I) wherein A is O, andwherein R₀, R₂ and R₃ are each hydrogen.

Alternatively, the compound of formula (XVIII) may be reacted withN-(phenylseleno)phthalimide (VI) or phenylselenyl chloride, in thepresence of a suitably selected a Lewis Acid such as borontrifluorideetherate, and the like, in a solvent such as DCM, 1,2-dichloroethane,and the like, at a temperature in the range of form about 0° C. to about50° C., to yield the corresponding cyclic ether of formula (XIX), whereX is Ph-Se.

The cyclic ether of formula (XIX) is then reacted with a suitablyselected reducing agent such as tri-n-butylstannane,tris(trimethylsilyl)silane, and the like, in the presence of a radicalinitiator such as AIBN, and the like, in a solvent such as benzene,toluene, and the like, at a temperature in the range of from about 60°C. to about 120° C. to yield the corresponding intermediate of formula(XX).

The intermediate of formula (XX) is then reacted as described above, toyield the corresponding compound of formula (I) wherein A is O, andwherein R₀, R₂ and R₃ are each hydrogen.

Additionally, reductive alkylation as described in Scheme 1 can beemployed by one of ordinary skill in the art as a means for transformingthese compounds of formula (I) where A is O and R₀ is H to compounds offormula (I) where A is O and R₀ is other than H.

EXAMPLES

Representative compounds of the present invention can be synthesized inaccordance with the general synthetic methods described below. Compoundsof Formula (I) can be prepared by methods known to those who are skilledin the art. The following examples are only meant to represent examplesof the invention and are in no way meant to be a limit of the invention.

Intermediate 1 (R)-3-methoxydihydro-2H-pyran-4(3H)-one

Step A 4,4-Dimethoxytetrahydro-2H-pyran-3-ol

A 12-L 4-neck round bottom flask with an overhead stirrer was chargedwith MeOH (8.18 L) and potassium hydroxide (400.5 g, 2.4 mol) whilestirring at room temperature until base was completely dissolved (anexotherm was observed). The homogeneous mixture was cooled to 0° C. withan ice-acetone bath. To a 500-mL addition funnel was charged withtetrahydro-4H-pyran-4-one (250 g, 2.5 mol) and after the KOH-methanolsolution temperature reached 0° C., the pyranone was added drop wisewhile maintaining temperature at <5° C. After stirring for an additional1.5 h, iodine (704 g, 1.1 mol) was added portion wise over a 1.5 hperiod while maintaining the temperature at <5° C. The reaction mixturewas allowed to stir at room temperature for 18 h. The reaction wasconcentrated and the remaining residue was treated with toluene (1.5 L)and stirred for ½ h. A solid had precipitated, was filtered off and thefiltrate was evaporated to afford to afford the title compound (330 g,81%) as an amber oil.

Step B 4,4-Dimethoxydihydro-2H-pyran-3(4H)-one

A 12-L 4-neck Morton flask equipped with an overhead stirrer,thermocouple, and two addition funnels was charged with oxalyl chloride(130 mL, 1.49 mol) and CH₂Cl₂ (2.5 L). The solution was chilled withdry-ice/acetone bath to −72° C. DMSO (178 mL, 2.50 mol) was added viaadditional funnel in CH₂Cl₂ (530 mL) over ½ h period while maintainingtemperature at or below −70° C. After the addition was complete, themixture was stirred for an additional 30 min and4,4-dimethoxytetrahydro-2H-pyran-3-ol (as prepared in Step A, 200 g,1.23 mol) in CH₂Cl₂ (630 mL) was added slowly (˜½ h) from an additionfunnel keeping the temperature at or below −70° C. After stirring anadditional 30 min, Et₃N (870 mL, 6.24 mol) was added, the temperaturereached −42° C. and dropped back down to approx. −70° C. The stirredmixture was allowed to stir to room temperature over 18 h. The mixturewas filtered and the filtrate was concentrated to provide the crudeproduct plus Et₃N—HCl solid. The mixture was filtered and rinsed withEtOAc (2×500 mL). The filtrate was concentrated again to a slurry. Theslurry was diluted with EtOAc (approx 1 L), filtered, and concentratedagain to give an amber oil containing the product and residual DMSO asthe primary components. After purification on silica gel using a mixtureof ethyl acetate in heptanes, the title compound (285 g, 90%) wasafforded as a brown solid.

Step C1 (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol

A 12-L 4-neck round bottom flask equipped with a overhead air stirrer,addition funnel with nitrogen inlet adapter, condenser, and thermocouplewas charged with S-(−)-2-methyl-CBS-oxazaborolidine (40 g, 0.12 mol) andTHF (2.2 L). The mixture was warmed under nitrogen to 40° C. thenMe₂S—BH₃ (108 mL, 1.15 mol) was added to the THF-catalyst mixture viasyringe. An addition funnel charged with4,4-dimethoxydihydro-2H-pyran-3(4H)-one (as prepared in the previousstep, 165 g, 0.59 mol) in THF (2.1 L) was added drop wise over a 7 hperiod. After the addition was complete, the reaction was allowed tostir for 18 h at 40° C. The reaction was chilled to 10° C. in anice-acetone bath and quenched by slow addition of MeOH (1.1 L) over 1 hperiod. The cooling bath was removed and the mixture allowed to warm toroom temperature for 3 h. After the gas evolution ceased, the mixturewas concentrated on a rotary evaporator to give 188 g. Purification onsilica gel using a mixture of EtOAc and heptanes afforded the titlecompound (166 g, 99%, chiral GC 93% ee) as a yellow oil.

Step C Alternative (R)-4,4-Dimethoxytetrahydro-2H-pyran-3-ol

A 12-L 4-neck round bottom flask equipped with a overhead air stirrer,addition funnel with nitrogen inlet adapter, condenser, and thermocouplewas charged with S-(−)-2-methyl-CBS-oxazaborolidine (78 g, 0.28 mol) andTHF (2.7 L). The mixture was warmed under nitrogen to 40° C. whileborane-N,N-diethylaniline complex (280 mL, 1.57 mol) was added to theTHF-catalyst mixture via addition funnel over 40 min. An 4-L Erlenmeyerflask was charged with a mixture of4,4-dimethoxydihydro-2H-pyran-3(4H)-one (as prepared in Step B, 225 g,1.4 mol) in THF (2.7 L) and was added drop wise over a 8 h period viametering pump. After addition, the reaction was allowed to stir for 18 hat 40° C. The reaction was chilled to 10° C. in an ice-acetone bath andquenched by slow addition of MeOH (1.35 L) over 1 h period; after MeOHaddition was complete, the cooling bath was removed and the mixtureallowed to warm to room temperature for 3 h. After the gas evolutionceased, the mixture was concentrated on a rotary evaporator to give 365g. After purification on silica gel using a mixture of EtOAc in heptanesthe title compound (157 g, 69%, chiral GC 95.5% ee) was afforded as ayellow oil.

Step D (R)-3,4,4-Trimethoxytetrahydro-2H-pyran

A 12-L 4-neck round bottom flask equipped with a overhead air stirrer,addition funnel with nitrogen inlet adapter, condenser, and thermocouplewas charged with (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol (as preparedin the previous step, 163 g, 1.0 mol) in THF (2.4 L) and stirred inice/acetone bath until <0° C. KOtBu (113 g, 1.0 mol) was added in oneportion, and after stirring 45 min, dimethyl sulfate (95 mL, 1.0 mol)was added via addition funnel over 15 min. The reaction was allowed tostir at room temperature for 2 h. The reaction mixture was poured into areparatory flask containing H₂O (1.2 L) and CH₂Cl₂ (1.2 L) and thelayers were separated. The aqueous layer was back-extracted with CH₂Cl₂(900 mL). The combined organic layers were washed with brine (1.0 L;Note: organic layer was on top), dried over MgSO₄, filtered andevaporated to afford the title compound (177.1 g, 99%, chiral GC 94.4%ee) as a light yellow oil.

Step E (R)-3-methoxydihydro-2H-pyran-4(3H)-one

A stirred mixture of (R)-3,4,4-trimethoxytetrahydro-2H-pyran (asprepared in the previous step, 141 g, 0.80 mol), in THF (3.6 L) and H₂O(1.1 L) in an ice/acetone bath at <0° C., was treated with a solution ofHCl (conc., 595 mL, 7.21 mol) added via addition funnel over 45 minwhile keeping temperature <3° C. After the addition was complete, thereaction was allowed to stir for 1.5 h at 0° C. The reaction wasevaporated until ˜1.9 L of concentrate remained. The concentrated wastransferred to a separatory funnel and extracted with CH₂Cl₂ (3×1 L).The combined organic fractions were washed with sat. NaHCO₃ (1 L), brine(1 L), dried over MgSO₄, filtered and evaporated to afford the titlecompound (71.4 g, 69%, Chiral GC 92% ee) as an oil which solidified uponstanding. Optical Rotation: [α]²⁵(D)-6.97° (c=0.8222, MeOH); ElementalAnalysis calc for C₆H₁₀O₃: C, 53.59; H, 7.58. Found: C, 53.64; H, 7.65.

Intermediate 2 (S)-3-methoxydihydro-2H-pyran-4(3H)-one

Step A (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol

A 5-L 4-neck round bottom flask equipped with an overhead air stirrer,addition funnel with nitrogen inlet adapter, condenser, and athermocouple was charged with (R)-(+)-2-methyl-CBS-oxazaborolidine (34g, 0.12 mol) and THF (1.2 L). The mixture was warmed under nitrogen to40° C. then Me₂S—BH₃ (63 mL, 0.67 mol) was added to the THF-catalystmixture via syringe. An addition funnel was charged with4,4-dimethoxydihydro-2H-pyran-3(4H)-one (as prepared in Intermediate 1,Step B, 96 g, 0.59 mol) in THF (1.2 L) was added drop wise over an 8 hperiod. After addition, the reaction was allowed to stir for 18 h at 40°C. The reaction was chilled to 10° C. in an ice-acetone bath andquenched by slow addition of MeOH (600 mL) over 45 min. The cooling bathwas removed and the mixture allowed to warm to room temperature for 3 h.After the gas evolution ceased, the mixture was concentrated on a rotaryevaporator to give 132 g. Purification on silica gel using a mixture ofEtOAc and heptanes afforded the title compound (80.5 g, 83%, chiral GC95% ee) as a yellow oil.

Step A Alternative 1 (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol

The reaction was carried out according to the procedure of Intermediate2, Step A, using 0.1 equivalents of(R)-(+)-2-methyl-CBS-oxazaborolidine, and substituting BH₃-THF complexfor Me₂S—BH₃. The product (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol wasobtained in 88% yield and 60% ee.

Step A Alternative 2 (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol

The reaction was carried out according to the procedure of Intermediate2, Step A, using 0.1 equivalents of(R)-(+)-2-methyl-CBS-oxazaborolidine, and substituting catecholboranefor Me₂S—BH₃. The product (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol wasobtained in <20% yield and 60% ee.

Step B (S)-3,4,4-trimethoxytetrahydro-2H-pyran

A 3-L 4-neck round bottom flask equipped with a overhead air stirrer,addition funnel with nitrogen inlet adapter, condenser, and thermocouplewas charged with (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol (as preparedin the previous step, 80 g, 0.49 mol) in THF (1.1 L) and stirred inice/acetone bath until <0° C. KOtBu (56 g, 0.49 mol) was added in oneportion, and after stirring 45 min, dimethyl sulfate (47 mL, 0.49 mol)was added via addition funnel over 15 min. The reaction was allowed tostir at room temperature for 3 h. The reaction mixture was poured into aseparatory flask containing H₂O (1.25 L) and CH₂Cl₂ (1.25 L) and thelayers were separated. The aqueous layer was back-extracted with CH₂Cl₂(750 mL). The combined organic layers were washed with brine (1 L; Note:organic layer was on top), dried over MgSO₄, filtered and evaporated toafford the title compound (83.5 g, 96%, chiral GC 94.6% ee) as a lightyellow oil.

Step C (S)-3-methoxydihydro-2H-pyran-4(3H)-one

A 5-L 4-neck round bottom flask equipped with a overhead air stirrer,nitrogen inlet adapter, thermocouple, and septum was charged with(S)-3,4,4-trimethoxytetrahydro-2H-pyran (as prepared in the previousstep, 83 g, 0.47 mol), THF (2.1 L), H₂O (670 mL), and stirred inice/acetone bath until <0° C., where upon a solution of HCl (conc., 350mL, 4.24 mol) was added via addition funnel over 30 min while keepingtemperature <2° C. After the addition was complete, the reaction wasallowed to stir for 1 h at 0° C. The reaction was evaporated until ˜1.2L of concentrate remained. The concentrate was transferred to areparatory funnel and extracted with CH₂Cl₂ (3×750 mL). The combinedorganic fractions were washed with sat. NaHCO₃ (500 mL), brine (500 mL),dried over MgSO₄, filtered and evaporated to afford the title compound(46.9 g, 77%, Chiral GC 91% ee) as an oil which solidified upon sitting.Optical Rotation: [α]²⁵(D)+3.65° (c=1.020, MeOH); Elemental Analysiscalc for C₆H₁₀O₃: C, 53.39; H, 7.57. Found: 53.59; H, 7.62.

Example 1((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step A (1S,4S)-methyl4-((tert-butoxycarbonyl)amino)-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)cyclopent-2-enecarboxylate

To a solution of LiHMDS in THF (72.9 mL of a 1 M solution, 72.9 mmol,2.2 eq) at −78° C. under Ar was added a solution of (1R,4S)-methyl4-((t-butoxycarbonyl)amino)cyclopent-2-enecarboxylate (preparedaccording to the procedure of US 20050101628 A1 (see page 31, column 1,Procedure B, step B)), 8.00 g, 33.2 mmol, 1 eq) in THF (40 mL) dropwiseover 1 hr. After stirring for 30 min, a solution oft-butyl(2-iodoethoxy)dimethylsilane (13.29 g, 46.4 mmol, 1.4 eq) in THF(20 mL) was added. The solution was kept at −78° C. for 15 min, thengradually warmed to 0° C. over 2 hrs, and kept at 0° C. for 1 hr. 1 NHCl and water were added, the solution extracted with DCM, the organicscombined, dried over MgSO₄, and concentrated. Purification bychromatography (400 g column) eluting with 5 to 20% EtOAc/heptaneafforded the title compound of Step A. ¹H NMR (CHLOROFORM-d) δ:5.76-5.85 (m, 2H), 4.90 (d, J=9.0 Hz, 1H), 4.72-4.82 (m, 1H), 3.69 (s,3H), 3.57-3.64 (m, 2H), 2.24 (dd, J=13.9, 8.0 Hz, 1H), 2.14 (dd, J=14.1,3.5 Hz, 2H), 1.71-1.82 (m, 1H), 1.40-1.50 (m, 9H), 0.84-0.90 (m, 9H),0.03 (s, 6H). ESI-MS (m/z): Calculated for C20H37NO5Si: 422.2 (M+23).found: 422.2.

Step B(1S,4S)-4-((tert-butoxycarbonyl)amino)-1-(2-((tert-butyldimethylsilyl)oxy)ethyl)cyclopent-2-enecarboxylicacid

To a solution of the product of Step A (7.89 g, 19.74 mmol, 1 eq) inmethanol (100 mL) at rt was added 1 N NaOH (59.2 mL, 59.2 mmol, 3.0 eq).After stirring overnight, the methanol was removed, 1 N HCl was addeduntil the solution was acidic, the solution extracted with DCM, theorganics combined, dried over MgSO₄, and concentrated to afford thetitle compound of Step B. ¹H NMR (CHLOROFORM-d) δ: 5.85 (br. s., 2H),4.97 (br. s., 1H), 4.80 (br. s., 1H), 3.71 (br. s., 2H), 1.99-2.41 (m,3H), 1.92 (br. s., 1H), 1.44 (br. s., 9H), 0.88 (s, 9H), 0.06 (br. s.,6H). ESI-MS (m/z): Calculated for C19H35NO5Si: 408.2 (M+23). found:408.3.

Step C tert-butyl((1S,4S)-4-(2-((tert-butyldimethylsilyl)oxy)ethyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of the product of Step B (3.47 g, 8.99 mmol, 1 eq) in DCM(40 mL) at rt was added HOBt hydrate (2.34 g, 15.3 mmol, 1.7 eq) andEDCI (2.58 g, 13.5 mmol, 1.5 eq). After 15 min, DIEA (7.8 mL, 45.3 mmol,5 eq) and 3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2HCl(3.71 g, 13.5 mmol, 1 eq) were added and the solution stirred overnightat rt. Saturated NaHCO₃ was added, the solution extracted with DCM, theorganics combined, dried over MgSO₄, and concentrated. Purification bychromatography (120 g column) eluting with 25 to 60% EtOAc/heptaneafforded the title compound of Step C. ¹H NMR (CHLOROFORM-d) δ: 8.71 (s,1H), 7.69 (s, 1H), 6.19 (d, J=5.4 Hz, 1H), 5.76 (dd, J=5.6, 2.0 Hz, 1H),4.68-4.86 (m, 4H), 3.85-4.07 (m, 2H), 3.54-3.65 (m, 2H), 3.13 (t, J=5.7Hz, 2H), 2.58 (dd, J=13.3, 7.7 Hz, 1H), 1.98-2.16 (m, 2H), 1.85-1.97 (m,1H), 1.42 (s, 9H), 0.84 (s, 9H), −0.02 (d, J=4.4 Hz, 6H). Calculated forC28H42F3N3O4Si: 570.3 (M+1). found: 570.3.

Step D tert-butyl((1S,4S)-4-(2-hydroxyethyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of the product of Step C (3.52 g, 6.18 mmol, 1 eq) in THF(50 mL) at rt was added TBAF in THF (12.36 mL of a 1 M solution, 12.36mmol, 2 eq). After 1 hr, water was added, the solution extracted withDCM, the organics combined, dried over MgSO₄, and concentrated.Purification by chromatography (80 g column) eluting with 2 to 6%methanol/DCM with ammonia afforded the title compound of Step D. ¹H NMR(CHLOROFORM-d) δ: 8.71 (s, 1H), 7.70 (s, 1H), 6.28 (dd, J=5.9, 2.0 Hz,1H), 5.78 (dd, J=5.9, 2.0 Hz, 1H), 4.70-4.95 (m, 4H), 3.99-4.10 (m, 1H),3.85-3.96 (m, 1H), 3.68 (br. s., 2H), 3.09-3.18 (m, 2H), 2.65 (dd,J=12.9, 7.4 Hz, 1H), 1.99-2.21 (m, 3H), 1.82-1.93 (m, 1H), 1.38-1.49 (m,9H). Calculated for C22H28F3N3O4: 456.2 (M+1). found: 456.2.

Step E tert-butyl((3aS,5S,6S,6aS)-6-(phenylselanyl)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)carbamate

To a solution of the product of Step D (1.51 g, 3.32 mmol, 1 eq) in DCM(40 mL) at rt under Ar was added N-(phenylseleno)phthalimide (1.60 g,4.97 mmol, 1.5 eq) and BF₃-etherate (0.042 mL, 0.33 mmol, 0.1 eq). After2 hrs, 1 N NaOH was added and stirred 5 min., water was added, thesolution extracted with DCM, the organics combined, dried over MgSO₄,and concentrated. Purification by chromatography (80 g column) elutingwith 50 to 100% EtOAc/heptane afforded the title compound of Step E. ¹HNMR (CHLOROFORM-d) δ: 8.72 (s, 1H), 7.69 (s, 1H), 7.50-7.62 (m, 2H),7.22-7.27 (m, 3H), 5.35 (s, 1H), 5.06 (d, J=8.1 Hz, 1H), 4.79-5.01 (m,1H), 4.66-4.78 (m, 1H), 4.53 (br. s., 1H), 3.78-4.06 (m, 4H), 3.68-3.78(m, 1H), 3.05-3.19 (m, 2H), 2.32 (dd, J=11.6, 5.8 Hz, 2H), 2.16 (d,J=10.9 Hz, 2H), 1.36 (s, 9H). Calculated for C28H32F3N3O4Se: 634.2(M+23). found: 634.1.

Step F tert-butyl((3aS,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)carbamate

To a solution of the product of Step E (1.51 g, 2.67 mmol, 1 eq),tris(trimethylsilyl)silane (1.72 mL, 5.34 mmol, 2 eq) and AIBN (438 mg,2.67 mmol, 1 eq) in benzene (20 mL) was warmed to 80° C. under Ar. After3 hrs, the solution was concentrated. Purification by chromatography (80g column) eluting with 50 to 100% EtOAc/heptane afforded the titlecompound of Step F. ¹H NMR (CHLOROFORM-d) δ: 8.72 (br. s., 1H), 7.71(br. s., 1H), 4.97-5.09 (m, 1H), 4.70-4.91 (m, 2H), 4.56-4.69 (m, 1H),4.28 (br. s., 1H), 3.80-4.07 (m, 3H), 3.71 (q, J=7.3 Hz, 1H), 3.13 (br.s., 2H), 2.07-2.53 (m, 4H), 1.81 (br. s., 1H), 1.61-1.72 (m, 1H), 1.40(s, 9H). Calculated for C22H28F3N3O4: 478.2 (M+23). found: 478.2.

Step G((3aS,5S,6aR)-5-aminohexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

To a solution of the product of Step F (12.54 g, 24.8 mmol, 1 eq) in DCM(100 mL) at rt was added TFA (20 mL, 261 mmol, 10.6 eq). After 1 hr, thesolution was concentrated. 3 M NaOH was added and the solution extractedwith DCM, the organics combined, dried over MgSO₄, and concentrated toafford the title compound of Step G. ¹H NMR (CHLOROFORM-d) δ: 8.72 (s,1H), 7.69 (br. s., 1H), 5.07 (d, J=4.9 Hz, 1H), 4.71-4.90 (m, 2H),3.84-4.03 (m, 3H), 3.58-3.71 (m, 2H), 3.09-3.20 (m, 2H), 2.14-2.41 (m,3H), 1.99-2.13 (m, 1H), 1.65-1.75 (m, 1H), 1.43-1.58 (m, 1H). Calculatedfor C17H20F3N302: 356.2 (M+1). found: 356.3.

Step H((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

To a solution of the product of Step G (119 mg, 0.33 mmol, 1 eq) in DCMat rt was added acetic acid (0.01 mL, 0.17 mmol, 0.5 eq),3-methoxytetrahydro-4H-pyran-4-one (131 mg, 1.0 mmol, 3 eq) and sodiumtriacetoxyborohydride (355 mg, 1.67 mmol, 5 eq). After stirringovernight, saturated NaHCO₃ was added, the solution extracted with DCM,the organics combined, dried over MgSO₄, and concentrated. Purificationby chromatography (12 g) eluting with 4 to 8% methanol/DCM with ammoniaafforded the title compound of Example 1. ¹H NMR (CHLOROFORM-d) δ: 8.72(br. s., 1H), 7.70 (br. s., 1H), 4.98-5.14 (m, 1H), 4.70-4.89 (m, 2H),3.80-4.18 (m, 5H), 3.25-3.75 (m, 8H), 3.07-3.24 (m, 2H), 2.53-2.89 (m,1H), 2.01-2.48 (m, 4H), 1.39-1.88 (m, 5H). Calculated for C23H30F3N3O4:470.2 (M+1). found: 470.2.

Separation of Example 1 by chiral HPLC gave 4 products, Example 2,Example 3, Example 4 and Example 5.

Example 2((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

¹H NMR (CHLOROFORM-d) δ: 8.71 (s, 1H), 7.70 (br. s., 1H), 5.05 (d, J=4.6Hz, 1H), 4.68-4.88 (m, 2H), 4.08 (dd, J=12.5, 2.9 Hz, 1H), 3.81-4.04 (m,4H), 3.66 (td, J=8.8, 6.8 Hz, 1H), 3.50-3.62 (m, 1H), 3.34-3.46 (m, 4H),3.24-3.34 (m, 2H), 3.14 (br. s., 2H), 2.76 (d, J=9.5 Hz, 1H), 2.14-2.46(m, 3H), 1.99-2.14 (m, 1H), 1.45-1.86 (m, 5H). Calculated forC23H30F3N3O4: 470.2 (M+1). found: 470.2.

Example 3((3aS,5S,6aR)-5-(((3R,4R)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

¹H NMR (CHLOROFORM-d) δ: 8.72 (s, 1H), 7.70 (br. s., 1H), 5.06 (d, J=4.6Hz, 1H), 4.78 (br. s., 2H), 4.05 (dd, J=12.6, 4.0 Hz, 1H), 3.84-4.02 (m,4H), 3.65 (td, J=8.9, 7.0 Hz, 1H), 3.55 (dt, J=9.7, 5.0 Hz, 1H),3.35-3.45 (m, 4H), 3.27-3.35 (m, 2H), 3.14 (br. s., 2H), 2.75-2.85 (m,1H), 2.35 (br. s., 1H), 2.24 (dd, J=13.0, 5.4 Hz, 2H), 2.00-2.15 (m,1H), 1.59-1.85 (m, 4H), 1.48 (ddd, J=13.2, 10.8, 4.9 Hz, 1H). Calculatedfor C23H30F3N3O4: 470.2 (M+1). found: 470.2.

Example 4((3aS,5S,6aR)-5-(((3S,4R)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone(the relative stereochemistry of the methoxypyran ring is trans, theabsolute stereochemistry is unknown but opposite that of Example 5)

¹H NMR (CHLOROFORM-d) δ: 8.72 (s, 1H), 7.70 (br. s., 1H), 5.01 (d, J=4.6Hz, 1H), 4.78 (br. s., 2H), 4.04-4.16 (m, 1H), 3.81-4.04 (m, 4H),3.49-3.70 (m, 2H), 3.27-3.44 (m, 4H), 3.15 (t, J=5.3 Hz, 2H), 2.96-3.09(m, 2H), 2.59 (br. s., 1H), 2.19-2.44 (m, 3H), 2.00-2.13 (m, 1H), 1.95(dt, J=13.4, 2.1 Hz, 1H), 1.62-1.87 (m, 2H), 1.57 (ddd, J=13.4, 10.9,5.0 Hz, 1H), 1.38-1.51 (m, 1H). Calculated for C23H30F3N3O4: 470.2(M+1). found: 470.2.

Example 5((3aS,5S,6aR)-5-(((3R,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone(the relative stereochemistry of the methoxypyran ring is trans, theabsolute stereochemistry is unknown but opposite that of Example 4)

¹H NMR (CHLOROFORM-d) δ: 8.72 (s, 1H), 7.69 (br. s., 1H), 5.06 (d, J=4.6Hz, 1H), 4.67-4.93 (m, 2H), 4.10 (d, J=7.1 Hz, 1H), 3.79-4.04 (m, 4H),3.48-3.69 (m, 2H), 3.29-3.42 (m, 4H), 3.14 (br. s., 2H), 2.96-3.10 (m,2H), 2.59-2.72 (m, 1H), 2.21-2.43 (m, 3H), 2.02-2.15 (m, 1H), 1.98 (d,J=24.5 Hz, 1H), 1.57-1.81 (m, 2H), 1.30-1.48 (m, 2H). Calculated forC23H30F3N3O4: 470.2 (M+1). found: 470.2.

Example 6((3aS,5S,6aR)-5-((tetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example1, Step G and tetrahydro-4H-pyran-4-one following the proceduredescribed in Example 1, Step H. ¹H NMR (CHLOROFORM-d) δ: 8.72 (br. s.,1H), 7.70 (br. s., 1H), 5.04 (d, J=4.6 Hz, 1H), 4.78 (br. s., 2H), 3.95(s, 2H), 3.97 (s, 3H), 3.53-3.75 (m, 2H), 3.28-3.49 (m, 2H), 3.14 (br.s., 2H), 2.62-2.81 (m, 1H), 2.29 (dd, J=12.8, 5.5 Hz, 3H), 1.99-2.17 (m,1H), 1.60-1.92 (m, 3H), 1.18-1.57 (m, 5H). Calculated for C22H28F3N3O3:440.2 (M+1). found: 440.2.

Example 7 ((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(7-(trifluoromethyl)-3,4-dihydroisoquinolin-2(1H)-yl)methanone

The title compound was prepared from reaction of the product of Step Bof Example 1 and 7-(trifluoromethyl)-1,2,3,4-tetrahydroisoquinolinefollowing the procedure described in Example 1, Step C, and thenfollowing Example 1 Steps D through H.

¹H NMR (CHLOROFORM-d) δ: 7.45 (d, J=7.6 Hz, 2H), 7.23-7.31 (m, 1H),4.99-5.10 (m, 1H), 4.72 (br. s., 2H), 4.02-4.16 (m, 1H), 3.70-4.02 (m,4H), 3.47-3.59 (m, 1H), 3.22-3.46 (m, 6H), 2.86-3.09 (m, 3H), 2.77 (br.s., 1H), 2.13-2.44 (m, 3H), 1.99-2.11 (m, 1H), 1.35-1.88 (m, 5H).Calculated for C24H31F3N2O4: 470.2 (M+1). found: 470.2.

Example 8((3aS,5S,6aR)-5-((tetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(7-(trifluoromethyl)-3,4-dihydroisoquinolin-2(1H)-yl)methanone

The title compound was prepared from reaction of the product of Step Bof Example 1 and 7-(trifluoromethyl)-1,2,3,4-tetrahydroisoquinolinefollowing the procedure described in Example 1, Step C, following StepsD through G, and then using tetrahydro-4H-pyran-4-one following theprocedure described in Example 1, Step H.

JNJ46713953, ¹H NMR (CHLOROFORM-d) δ: 7.31-7.50 (m, 2H), 7.26-7.29 (m,1H), 5.05 (d, J=4.6 Hz, 1H), 4.72 (br. s., 2H), 3.90-4.05 (m, 3H),3.70-3.85 (m, 2H), 3.52-3.70 (m, 2H), 3.30-3.45 (m, 2H), 2.95 (br. s.,2H), 2.70 (br. s., 1H), 2.18-2.44 (m, 3H), 2.00-2.12 (m, 1H), 1.60-1.89(m, 2H), 1.23-1.53 (m, 5H). Calculated for C23H29F3N2O3: 439.2 (M+1).found: 439.2.

Example 9 ((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(7-(trifluoromethoxy)-3,4-dihydroisoquinolin-2(1H)-yl)methanone

The title compound was prepared from reaction of the product of Step Bof Example 1 and 7-(trifluoromethoxy)-1,2,3,4-tetrahydroisoquinolinefollowing the procedure described in Example 1, Step C, and thenfollowing Example 1 Steps D through H. Calculated for C24H31F3N2O5:485.2 (M+1). found: 485.2.

Example 10((3aS,5S,6aR)-2-cyclopropyl-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step A tert-butyl((1S,4S)-4-(2-oxoethyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of the product of Example 1, Step D (417 mg, 0.92 mmol, 1eq) in DCM (20 mL) at 0° C. was added Dess-Martin periodinane (427 mg,1.01 mmol, 1.1 eq). After 1 hr, saturated sodium bicarbonate and sodiumthiosulfate were added, after 10 minutes, the aqueous was extracted withDCM, the organics combined, dried over MgSO₄ and concentrated.Purification by chromatography (12 g column) eluting with 30 to 60%EtOAc/heptane afforded the title compound of Step A. ¹H NMR(CHLOROFORM-d) δ: 9.74 (s, 1H), 8.70 (s, 1H), 7.72 (s, 1H), 6.31 (dd,J=5.6, 1.7 Hz, 1H), 5.91 (dd, J=5.6, 1.5 Hz, 1H), 5.01 (d, J=17.1 Hz,1H), 4.65-4.87 (m, 3H), 4.07-4.21 (m, 1H), 3.79-3.95 (m, 1H), 3.10 (q,J=5.6 Hz, 2H), 3.03 (d, J=16.6 Hz, 1H), 2.63 (dd, J=13.6, 7.2 Hz, 1H),2.52 (dd, J=16.6, 1.5 Hz, 1H), 2.11 (dd, J=13.1, 7.7 Hz, 1H), 1.44 (s,9H). Calculated for C22H26F3N3O4: 454.2 (M+1). found: 454.2.

Step B tert-butyl((1S,4S)-4-(2-cyclopropyl-2-hydroxyethyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of cyclopropyl magnesium bromide (6.88 mL of a 0.5 Msolution in THF, 3.44 mmol, 8 eq) in THF (5 mL) at 0° C. under Ar wasadded a solution of the product of Step A (195 mg, 0.43 mmol, 1 eq) inTHF (17 mL) dropwise over 30 min. After 45 min, saturated NH₄Cl wasadded, the solution extracted with ethyl acetate, the organics combined,dried over MgSO₄ and concentrated. Purification by chromatography (12 gcolumn) eluting with 30 to 100% EtOAc/heptane afforded the titlecompound of Step B as a mix of diastereomers. ¹H NMR (CHLOROFORM-d) δ:8.70 (br. s., 1H), 7.70 (br. s., 1H), 6.11-6.42 (m, 1H), 5.77 (t, J=5.1Hz, 1H), 4.81 (d, J=12.2 Hz, 4H), 4.03 (dt, J=12.8, 6.2 Hz, 1H),3.81-3.98 (m, 1H), 3.59-3.75 (m, 2H), 3.03-3.25 (m, 2H), 2.69-3.00 (m,2H), 2.19-2.67 (m, 3H), 1.91-2.18 (m, 3H), 1.55-1.88 (m, 5H), 1.42 (s,9H), 0.88-0.99 (m, 1H), 0.42-0.56 (m, 2H), 0.14-0.35 (m, 2H). Calculatedfor C25H32F3N3O4: 496.2 (M+1). found: 496.2.

The title compound of Example 10 was made by taking the product ofExample 10, Step B and following the procedures from Example 1, Steps Ethrough H.

¹H NMR (CHLOROFORM-d) δ: 8.72 (s, 1H), 7.69 (br. s., 1H), 5.01-5.29 (m,1H), 4.65-4.90 (m, 2H), 3.74-4.15 (m, 5H), 3.70 (dd, J=11.0, 2.9 Hz,1H), 3.23-3.55 (m, 7H), 3.14 (br. s., 2H), 3.05 (ddd, J=10.0, 8.2, 6.0Hz, 1H), 2.69-2.87 (m, 1H), 2.36-2.52 (m, 1H), 2.06-2.36 (m, 3H),1.66-1.97 (m, 2H), 1.33-1.52 (m, 1H), 0.83-0.98 (m, 1H), 0.43-0.66 (m,2H), 0.36 (dt, J=8.6, 4.1 Hz, 1H), 0.10-0.24 (m, 1H). Calculated forC26H34F3N3O4: 510.3 (M+1). found: 510.3.

Example 11((3aS,5S,6aR)-2-ethyl-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and ethyl magnesium bromide following the procedure describedin Example 10, Step B, and then taking that product and following theprocedures from Example 1, Steps E through H. ¹H NMR (MeOD) δ: 8.72 (s,1H), 8.05 (br. s., 1H), 4.94-5.13 (m, 1H), 4.70-4.86 (m, 2H), 4.25 (br.s., 1H), 3.76-4.14 (m, 4H), 3.62-3.75 (m, 1H), 3.35-3.60 (m, 7H), 3.13(dd, J=3.3, 1.6 Hz, 2H), 2.33-2.75 (m, 3H), 1.69-2.10 (m, 5H), 1.39-1.69(m, 2H), 0.87-1.01 (m, 3H). Calculated for C25H34F3N3O4: 498.3 (M+1).found: 498.2.

Example 12((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2-methylhexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and methyl magnesium bromide following the proceduredescribed in Example 10, Step B, and then taking that product andfollowing the procedures from Example 1, Steps E through H. ¹H NMR(MeOD) δ: 8.72 (br. s., 1H), 8.05 (br. s., 1H), 4.96-5.21 (m, 1H),4.70-4.87 (m, 2H), 4.21-4.36 (m, 1H), 3.77-4.18 (m, 5H), 3.32-3.62 (m,7H), 3.00-3.23 (m, 2H), 2.32-2.76 (m, 3H), 1.68-2.06 (m, 5H), 1.16-1.29(m, 3H). Calculated for C24H32F3N3O4: 484.2 (M+1). found: 484.2.

Example 13((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2-propylhexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and propyl magnesium bromide following the proceduredescribed in Example 10, Step B, and then taking that product andfollowing the procedures from Example 1, Steps E through H. Calculatedfor C26H36F3N3O4: 512.3 (M+1). found: 512.2.

Example 14((3aS,5S,6aR)-2-isobutyl-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and isobutyl magnesium bromide following the proceduredescribed in Example 10, Step B, and then taking that product andfollowing the procedures from Example 1, Steps E through H.

Calculated for C27H38F3N3O4: 526.3 (M+1). found: 526.3.

Example 15((3aS,5S,6aR)-2-cyclohexyl-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and cyclohexyl magnesium bromide following the proceduredescribed in Example 10, Step B, and then taking that product andfollowing the procedures from Example 1, Steps E through H.

¹H NMR (CHLOROFORM-d) δ: 8.71 (br. s., 1H), 7.69 (br. s., 1H), 4.98-5.21(m, 1H), 4.65-4.94 (m, 2H), 3.68-4.17 (m, 5H), 3.23-3.64 (m, 8H), 3.13(br. s., 2H), 2.69-2.86 (m, 1H), 1.86-2.35 (m, 6H), 1.32-1.79 (m, 8H),1.10-1.30 (m, 3H), 0.86-1.06 (m, 2H). Calculated for C29H40F3N3O4: 552.3(M+1). found: 553.2.

Example 16((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2-phenylhexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and phenyl magnesium bromide following the proceduredescribed in Example 10, Step B, and then taking that product andfollowing the procedures from Example 1, Steps E through H. Calculatedfor C29H34F3N3O4: 546.3 (M+1). found: 546.3.

Example 17((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2-(1-methyl-1H-pyrazol-5-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step A tert-butyl((1S,4S)-4-(2-hydroxy-2-(1-methyl-1H-pyrazol-5-yl)ethyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of 1-methylpyrazole (0.165 mmol, 1.98 mmol, 3 eq) in THF(8 mL) at −78° C. under Ar was added n-BuLi (0.77 mL of a 2.5 M solutionin hexane, 1.92 mmol, 2.9 eq) and the solution stirred 1 hr. Then asolution of the product of Example 10, Step A (300 mg, 0.66 mmol, 1 eq)in THF (8 mL) was added over 5 min. After 1 hr, saturated NH₄Cl wasadded, the solution extracted with ethyl acetate, the organics combined,dried over MgSO₄ and concentrated. Purification by chromatography (12 gcolumn) eluting with 50 to 100% EtOAc/heptane to 5 to 10% methanol/DCMafforded the title compound of Step A as a mix of diastereomers. ¹H NMR(CHLOROFORM-d) δ: 8.70 (s, 1H), 7.62-7.80 (m, 1H), 7.33 (d, J=8.8 Hz,1H), 6.19-6.49 (m, 1H), 6.09 (br. s., 1H), 5.87 (dd, J=17.9, 5.6 Hz,1H), 4.54-5.03 (m, 5H), 3.74-3.96 (m, 4H), 3.48 (s, 1H), 2.90-3.23 (m,2H), 2.46-2.87 (m, 2H), 2.06-2.39 (m, 2H), 1.73-1.98 (m, 1H), 1.43 (br.s., 9H). Calculated for C27H34F3N5O4: 558.2 (M+23). found: 558.2.

The title compound of Example 17 was made by taking the product ofExample 17, Step A and following the procedures from Example 1, Steps Ethrough H.

¹H NMR (CHLOROFORM-d) δ: 8.73 (br. s., 1H), 7.72 (br. s., 1H), 7.32-7.48(m, 1H), 5.90-6.28 (m, 1H), 5.08-5.36 (m, 1H), 4.64-4.98 (m, 2H),3.76-4.17 (m, 7H), 3.50-3.74 (m, 1H), 3.23-3.50 (m, 6H), 3.16 (br. s.,2H), 2.66-2.86 (m, 1H), 2.15-2.64 (m, 4H), 1.96-2.05 (m, 1H), 1.42-1.91(m, 3H). Calculated for C27H34F3N5O4: 550.3 (M+1). found: 550.2.

Example 18 ((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2-(1-methyl-1H-imidazol-2-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and 1-methylimidazole following the procedure described inExample 17, Step A, and then taking that product and following theprocedures described in Example 1, Steps E through H.

Calculated for C27H34F3N5O4: 550.3 (M+1). found: 550.2.

Example 19((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2-(thiazol-2-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example10, Step A and thiazole following the procedure described in Example 17,Step A, and then taking that product and following the proceduresdescribed in Example 1, Steps E through H. Calculated for C26H31F3N4O4S:553.3 (M+1). found: 553.3.

Example 20 ((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-2,2-dimethylhexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step A2-((1S,4S)-4-((tert-butoxycarbonyl)amino)-1-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)aceticacid

To a solution of the product of Example 1, Step D (508 mg, 1.12 mmol, 1eq) in acetone (10 mL) at 0° C. was added Jones oxidation solution (0.46mL, 1.23 mmol, 1.1 eq). After 2 hr, water was added, the aqueous wasextracted with ethyl acetate, the organics combined, dried over MgSO₄and concentrated to afford the product of Step A that was usedunpurified in the next step. Calculated for C22H26F3N3O5: 492.2 (M+23).found: 492.1.

Step B methyl2-((1S,4S)-4-((tert-butoxycarbonyl)amino)-1-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)acetate

To a solution of the product of Step A (436 mg, 0.84 mmol, 1 eq) inmethanol (20 mL) at 0° C. was added trimethylsilyl diazomethane (5 mL ofa 2 M solution in hexanes, 10 mmol, 11.9 eq) until the yellow colorpersisted. The yellow solution was concentrated. Purification bychromatography (24 g column) eluting with 40 to 80% EtOAc/heptaneafforded the product of Step B. ¹H NMR (CHLOROFORM-d) δ: 8.70 (s, 1H),7.69 (s, 1H), 6.37 (d, J=5.4 Hz, 1H), 5.87 (d, J=5.4 Hz, 1H), 4.98 (d,J=17.4 Hz, 1H), 4.63-4.83 (m, 3H), 4.07-4.20 (m, 1H), 3.79-3.93 (m, 1H),3.64 (s, 3H), 3.08-3.19 (m, 2H), 3.04 (d, J=15.9 Hz, 1H), 2.62 (dd,J=13.4, 7.1 Hz, 1H), 2.46 (d, J=15.7 Hz, 1H), 2.01-2.12 (m, 1H), 1.43(s, 9H). Calculated for C23H28F3N3O5: 506.2 (M+23). found: 506.2.

Step C tert-butyl((1S,4S)-4-(2-hydroxy-2-methylpropyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of methyl magnesium chloride (2.62 mL of a 3 M solution inTHF, 7.86 mmol, 20 eq) in THF (6 mL) at 0° C. under Ar was added asolution of the product of Step B (190 mg, 0.39 mmol, 1 eq) in THF (6mL) dropwise over 30 min. After 30 min, saturated NH₄Cl was added, thesolution extracted with ethyl acetate, the organics combined, dried overMgSO₄ and concentrated. Purification by chromatography (12 g column)eluting with 40 to 100% EtOAc/heptane afforded the product of Step C. ¹HNMR (CHLOROFORM-d) δ: 8.70 (s, 1H), 7.69 (s, 1H), 6.47 (d, J=5.4 Hz,1H), 5.75 (dd, J=5.6, 1.5 Hz, 1H), 4.60-4.96 (m, 4H), 4.05 (d, J=13.7Hz, 1H), 3.89 (dt, J=13.3, 6.4 Hz, 1H), 3.07-3.19 (m, 2H), 2.54 (dd,J=12.6, 6.5 Hz, 1H), 2.10-2.27 (m, 2H), 1.80-1.96 (m, 2H), 1.35-1.49 (m,9H), 1.26 (s, 3H), 1.22 (s, 3H). Calculated for C24H32F3N3O4: 506.2(M+23). found: 506.2.

The title compound of Example 20 was made by taking the product of StepC and following the procedures described in Example 1, Steps E throughH.

¹H NMR (CHLOROFORM-d) δ: 8.72 (br. s., 1H), 7.69 (br. s., 1H), 5.16-5.27(m, 1H), 4.78 (br. s., 2H), 4.01-4.13 (m, 1H), 3.75-3.99 (m, 4H),3.53-3.67 (m, 1H), 3.22-3.48 (m, 6H), 3.12 (br. s., 2H), 2.79 (d, J=10.0Hz, 1H), 2.09-2.40 (m, 3H), 1.84-2.00 (m, 2H), 1.68-1.78 (m, 2H),1.42-1.53 (m, 1H), 1.34 (s, 3H), 1.16 (s, 3H). Calculated forC25H34F3N3O4: 498.2 (M+1). found: 498.2.

Example 21(3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-2-one

The title compound of Example 21 was made by taking the product ofExample 20, Step A and following the procedures described in Example 1,Steps E through H.

¹H NMR (CHLOROFORM-d) δ: 8.74 (br. s., 1H), 7.71 (br. s., 1H), 5.70 (d,J=4.2 Hz, 1H), 4.55-4.88 (m, 2H), 4.08 (d, J=11.7 Hz, 1H), 3.93 (d,J=11.5 Hz, 1H), 3.80 (br. s., 1H), 3.47-3.61 (m, 2H), 3.24-3.46 (m, 5H),3.15 (br. s., 2H), 3.03 (d, J=14.2 Hz, 1H), 2.83 (br. s., 2H), 2.23-2.47(m, 2H), 1.53-1.76 (m, 6H). Calculated for C23H28F3N3O5: 484.2 (M+1).found: 484.2.

Example 22((3aR,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]thiophen-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step AS-(2-((1S,4S)-4-((tert-butoxycarbonyl)amino)-1-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)ethyl)ethanethioate

To a solution of the product of Example 1, Step D (1950 mg, 3.98 mmol, 1eq) in THF (40 mL) at rt under Ar was added triphenylphosphine (2.09 g,7.96 mmol, 2 eq), diisopropylazodicarboxylate (1.55 mL, 7.96 mmol, 2 eq)and thioacetic acid (0.59 mL, 7.96 mmol, 2 eq). After 2 hr, water andsaturated NaHCO₃ were added, the aqueous was extracted with ether, theorganics combined, dried over MgSO₄ and concentrated. Purification bychromatography (80 g column) eluting with 30 to 60% EtOAc/heptaneafforded the product of Step A. ¹H NMR (CHLOROFORM-d) δ: 8.71 (s, 1H),7.71 (s, 1H), 6.21 (dd, J=5.6, 1.7 Hz, 1H), 5.80 (dd, J=5.6, 2.0 Hz,1H), 4.75-4.94 (m, 3H), 4.64-4.75 (m, 1H), 3.96-4.07 (m, 1H), 3.86-3.96(m, 1H), 3.14 (t, J=5.7 Hz, 2H), 2.69-2.79 (m, 2H), 2.65 (dd, J=13.4,8.1 Hz, 1H), 2.27 (s, 3H), 1.95-2.13 (m, 2H), 1.81-1.94 (m, 1H), 1.44(s, 9H). Calculated for C24H30F3N3O4S: 536.2 (M+23). found: 536.2.

Step B tert-butyl((1S,4S)-4-(2-mercaptoethyl)-4-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)cyclopent-2-en-1-yl)carbamate

To a solution of the product of Step A (1.74 g, 3.39 mmol, 1 eq) inmethanol (100 mL) at rt under Ar (degassed) was added 0.2 N NaOH (85 mL,85 mmol, 5 eq) that was degassed by bubbling Ar through the solutionprior to addition. After 2 hr, the methanol was concentrated, 6 N HClwas added until the solution was acidic, the aqueous was extracted withDCM, the organics combined, dried over MgSO₄ and concentrated to affordthe product of Step B which was used unpurified in the next step.Calculated for C22H28F3N3O3S: 494.2 (M+23). found: 494.1.

Step C tert-butyl((3aR,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]thiophen-5-yl)carbamate

To a solution of the product of Step B (1.25 g, 2.65 mmol, 1 eq) inbenzene (300 mL) at rt under Ar (degassed) was added AIBN (435 mg, 2.65mmol, 1 eq) and the solution heated to 85° C. for 3 days, thenconcentrated. Purification by chromatography (40 g column) eluting with25 to 60 to 100% EtOAc/heptane afforded the product of Step C. ¹H NMR(CHLOROFORM-d) δ: 8.72 (s, 1H), 7.69 (s, 1H), 4.65-4.90 (m, 3H),4.23-4.60 (m, 2H), 3.87-4.06 (m, 2H), 3.10-3.16 (m, 2H), 2.99-3.09 (m,1H), 2.87-2.98 (m, 1H), 1.95-2.38 (m, 6H), 1.40 (s, 9H). Calculated forC22H28F3N3O3S: 494.2 (M+23). found: 494.1.

The title compound of Example 22 was made by taking the product ofExample 22, Step C and following the procedures described in Example 1,Steps G and H.

¹H NMR (CHLOROFORM-d) δ: 8.71 (s, 1H), 7.69 (s, 1H), 4.73-4.93 (m, 2H),4.67 (br. s., 1H), 4.04-4.17 (m, 1H), 3.94 (t, J=5.9 Hz, 3H), 3.61-3.77(m, 1H), 3.36-3.47 (m, 4H), 3.26-3.36 (m, 2H), 3.06-3.19 (m, 2H),2.80-3.05 (m, 3H), 2.31 (br. s., 2H), 2.05-2.21 (m, 3H), 1.83-1.99 (m,1H), 1.60-1.83 (m, 2H). Calculated for C23H30F3N3O3S: 486.2 (M+1).found: 486.2.

Example 23((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone

Step A tert-butyl ((5S,7S)-1-oxo-2-oxaspiro[4.4]non-8-en-7-yl)carbamate

A 12-L three-neck round bottom flask with overhead mechanical stirrer,2-L addition funnel and nitrogen inlet was charged the product ofExample 1, Step A (1149 g, 2.875 mol, 1 eq) and THF (5.75 L). Theaddition funnel was charged with TBAF (1M solution in THF, 2.875 L,2.875 mol, 1 eq) and this solution was added dropwise over ˜1 h. Thetemperature increased from 17 to 21° C. and the reaction was clearorange at the end. The reaction was stirred for 1 h at rt, when it wasjudged complete by TLC and HPLC. The reaction was poured into a 22-Lseparatory flask charged with EtOAc (4 L) and the organic layer waswashed with brine (2 L). The organic layer was washed with additionalbrine (3×2 L) and these aqueous fractions were discarded. Heptane (4 L)was added and the organic layer was washed with water (3×2 L), brine(2×2 L) and the clear organic layer was checked by NMR for removal ofn-Bu₄NX. The organic layer was evaporated at 45° C. to about 750 mL,when the solution became hazy, heptane (800 mL) was added and instantcrystallization of a white solid resulted. More heptane (300 mL) wasadded and the mixture was swirled at 40° C. for 10 min on the rotovapbath. Ice was added to bath and the suspension was stirred at 13° C. for10 min. The solid was filtered on a Buchner funnel, washed with heptane(3×100 mL) and provided the product of Step A. ¹H NMR (CHLOROFORM-d) δ:6.02 (dd, J=5.4, 2.4 Hz, 1H), 5.77 (d, J=5.4 Hz, 1H), 5.21 (d, J=9.0 Hz,1H), 4.91 (t, J=8.7 Hz, 1H), 4.37 (dd, J=7.5, 6.5 Hz, 2H), 2.21-2.39 (m,3H), 2.07 (dd, J=13.8, 2.6 Hz, 1H), 1.44 (s, 9H). Calculated forC13H19NO4: 276.1 (M+23). found: 276.1.

Step B tert-butyl((1S,4S)-4-(2-hydroxyethyl)-4-(hydroxymethyl)cyclopent-2-en-1-yl)carbamate

To a 5-L three-neck round bottom flask equipped with a mechanicalstirrer, Claisen adapter with temperature probe and nitrogen inlet, andN₂ outlet was purged with nitrogen for 2 h before use. The product ofStep A (255.9 g, 1.01 mol, 1 eq) and MeOH (2 L) were added and thesolution was chilled to 2° C. in an ice bath. NaBH₄ (75 g, 1.98 mol, 2eq) was added in ˜5 equal portions; the temperature exothermed to 17° C.before coming back to 6° C., when the next portion was added. Theaddition took ˜2.5 h and the reaction was judged complete by HPLC afterthe last addition. The reaction was quenched at 7° C. by addition ofaqueous NH₄Cl (saturated, 1 L), wherein the temperature rose to about10° C. The white hazy mixture was concentrated on a rotary evaporate toabout 1 L (45° C. bath) when a white solid with some liquid resulted.The mixture was diluted with water and EtOAc (1 L each), transferred toa separatory funnel and the layers were separated. The aqueous layer wasextracted with EtOAc (3×250 mL). The combined organics were washed withbrine (125 mL), dried over MgSO₄, filtered through Celite and evaporatedat a bath temperature of 55° C. and provided the product of Step B as athick oil. ¹H NMR (CHLOROFORM-d) δ: 5.74 (s, 2H), 4.65-4.95 (m, 2H),3.70 (t, J=5.7 Hz, 2H), 3.40-3.57 (m, 2H), 2.76 (br. s., 1H), 2.28 (br.s., 1H), 2.19 (dd, J=13.4, 8.8 Hz, 1H), 1.70 (t, J=5.7 Hz, 2H), 1.55(dd, J=13.8, 4.0 Hz, 1H), 1.44 (s, 9H). Calculated for C13H23NO4: 280.2(M+23). found: 280.2.

Step C tert-butyl ((3aR,5S,6S,6aS)-3a-(hydroxymethyl)-6-(phenylselanyl)hexahydro-2H-cyclopenta[b]furan-5-yl)carbamate

To a 12-L three-neck round bottom flask equipped with a mechanicalstirrer, Claisen adapter with a nitrogen inlet and a temperature probe,and a nitrogen outlet was charged with the product of Step B (382 g,1.26 mol, 1 eq) and CH₂Cl₂ (6.5 L). N-(phenylseleno)phthalimide (419 g,1.39 mol, 1.1 eq) was added followed by BF₃ etherate (16 mL, 0.126 mol,0.1 eq) directly added by graduated cylinder. The reaction steadilyclimbed from 15° C. to 24° C. and within 10 min, the reaction formed apink precipitate. Ten min later, the reaction became thick with a whiteprecipitate and the temperature began to decrease. The reaction waschecked by HPLC and found to be complete. The reaction was filteredthrough Celite (removing the phthalimide impurity), the filter cake waswashed with CH₂Cl₂ (750 mL) until the filtrate was no longer orange. Thefiltrate was transferred to a separatory funnel, washed with aqueousNaOH (0.5 M, 2×1350 mL), brine (2×1 L), and the organic layer was driedover Na₂SO₄. [A second run was conducted with 382 g of the product ofStep B, under the same conditions, and was worked up and combined atthis point]. With about 3 L organics left, toluene (3 L, ˜4 mL/gstarting material) was added and the evaporation continued. Shortlyafter the toluene addition, crystallization occurred. The 20-L roundbottom flask was transferred to a heating mantel, and the contentsheated to 80° C., until the solid dissolved. The flask was transferredback to the rotary evaporator, reference material was used to seed thecrystallization, and the flask swirled (no heat) until the productstarted to crystallize. Ice was added to the bath and the contents ofthe flask swirled at 15° C. (external temp) for 30 min. The product wasfiltered, washed with ice-cold toluene and air-dried for 1 h, andafforded the product of Step C. ¹H NMR (CHLOROFORM-d) δ: 7.49-7.56 (m,2H), 7.23-7.29 (m, 3H), 5.05 (br. s., 1H), 4.46 (br. s., 1H), 4.29 (s,1H), 3.86-3.97 (m, 2H), 3.57-3.69 (m, 3H), 1.98-2.08 (m, 1H), 1.86-1.97(m, 2H), 1.73-1.86 (m, 2H), 1.41 (s, 9H). Calculated for C19H27NO4Se:436.1 (M+23). found: 436.1.

Step D tert-butyl((3aR,5S,6aR)-3a-(hydroxymethyl)hexahydro-2H-cyclopenta[b]furan-5-yl)carbamate

A 22-L four-neck round bottom flask equipped with mechanical stirrer,heating mantel, temperature probe, nitrogen inlet, and a refluxcondenser with nitrogen outlet was purged with nitrogen for 30 minbefore use. The product of Step C (603.5 g, 1.46 mol, 1 eq), AIBN (241g, 1.46 mol, 1 eq), tris(trimethylsilyl)silane (910 mL, 2.93 mol, 2 eq)and toluene (16.3 L) were added and the suspension was degassed withnitrogen purge through the suspension for 20 min. The reaction washeated to 80-83° C. for 1 h after which time the heat was shut off, andthe reaction cooled to rt over 12-18 h. TLC showed the reaction wascomplete. The reaction was poured directly into a BIOTAGE dry 5-kgcolumn that was eluted with 16 L of 50% EtOAc in heptane, followed by 32L of EtOAc and provided the product of Step D of a golden thick oil,which slowly crystallized on standing. ¹H NMR (CHLOROFORM-d) δ: 4.64(brd. s, 1H), 4.07-4.25 (m, 2H), 3.89 (ddd, J=8.8, 7.2, 4.5 Hz, 1H),3.53-3.67 (m, 3H), 2.17 (dd, J=13.3, 6.2 Hz, 1H), 1.98-2.06 (m, 1H),1.86-1.97 (m, 1H), 1.68-1.81 (m, 2H), 1.46-1.56 (m, 2H), 1.44 (s, 9H).Calculated for C13H23NO4: 202.2 (M−55). found: 202.2.

Step E(3aS,5S,6aR)-5-((tert-butoxycarbonyl)amino)hexahydro-2H-cyclopenta[b]furan-3a-carboxylicacid

A 22-L four-neck round bottom flask equipped with a mechanical stirrer,nitrogen inlet, 1-L addition funnel with nitrogen outlet, a temperatureprobe, and an external bath for cooling was purged with nitrogenovernight. A solution of the product of Step D (426 g, 1.57 mol) andacetone (8.1 L) was added, the flask was cooled to 7° C., and theaddition funnel was charged with Jones reagent (710 mL). The oxidant wasadded dropwise over 1 h 20 min, keeping the temp between 7-9° C. Afterthe first 200 mL was added, a green ball formed that made stirring verydifficult. After about ½ of the oxidant was added, LCMS was run tofollow the reaction. At the end of addition, olive green suspensionresulted with a hint of red (excess Jones). The ice bath was removed,the reaction was stirred at rt for 1 h after which time the reaction wasjudged complete. Isopropyl alcohol (40 mL) was added, the reactionstirred for 25 min, and water (800 mL) was added, that caused a niceseparation of green chunk from the acetone/water layer. Thewater/acetone was decanted off and evaporated. The green chunk wasdissolved in water (1.5 L), transferred to a separatory funnel andextracted with CH₂Cl₂ (1 L). The aqueous layer was checked by TLC andfound to contain no product, so it was discarded. The organic extractwas saved for combining later. The green water/acetone concentrate wasevaporated to about 5-7 L, until the solution looked hazy. Theconcentrate was transferred to a separatory funnel and extracted withCH₂Cl₂ (1×3 L, 3×1 L) and the aqueous layer checked after eachextraction for the presence of product. The combined extracts werewashed with brine (250 mL) which caused a terrible emulsion. Theemulsion was broken by addition of water and EtOAc (˜500 mL). Theorganic layer was dried (Na₂SO₄) but not very effectively as some watercame through during the filtration. Near the end of the evaporation, thedistillation rate slowed, and a thick yellow oil resulted. MeCN (500 mL)was added to the pot, the rotary evaporator bath warmed to 50° C., andthe contents seeded with reference material. A fine white solid slowlyformed within 10 min or so. Seeding was done a second time, andcontinued swirling for another 10 min at 50° C. Crystallization wasvisually detected, the bath was drained and filled with ice, and theflask swirled at 0° C. for 30 min, resulting in a thick white solid. Thesolid was filtered, washed with ice-cold MeCN (2×100 mL) and the solidwas air-dried overnight. The product of Step E was isolated as a white,free-flowing solid. ¹H NMR (MeOH) δ: 4.43 (d, J=5.4 Hz, 1H), 4.00-4.13(m, 1H), 3.89-3.98 (m, 1H), 3.63 (td, J=9.1, 5.7 Hz, 1H), 2.54 (ddd,J=12.6, 5.7, 3.2 Hz, 1H), 1.93-2.13 (m, 3H), 1.74-1.87 (m, 1H),1.53-1.66 (m, 1H), 1.43 (s, 9H). Calculated for C13H21NO5: 294.1 (M+23).found: 294.1.

Step F (benzyl4-((3aS,5S,6aR)-5-((tert-butoxycarbonyl)amino)hexahydro-2H-cyclopenta[b]furan-3a-carbonyl)piperazine-1-carboxylate

The product of Step F was prepared from the reaction of the product ofStep E and benzyl piperazine-1-carboxylate following the procedure fromExample 1, Step C. Calculated for C25H35N3O6: 496.2 (M+23). found:496.0.

Step G benzyl4-((3aS,5S,6aR)-5-aminohexahydro-2H-cyclopenta[b]furan-3a-carbonyl)piperazine-1-carboxylate

The product of Step G was prepared from the reaction of the product ofStep F following the procedure from Example 1, Step G. Calculated forC20H27N3O4: 374.2 (M+1). found: 374.2.

Step H benzyl4-((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-carbonyl)piperazine-1-carboxylate

The product of Step H was prepared from the reaction of the product ofStep G and (R)-3-methoxydihydro-2H-pyran-4(3H)-one (Intermediate 1)following the procedure described in Example 1, Step H. Calculated forC26H37N3O6: 488.3 (M+1). found: 488.1.

Step I((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(piperazin-1-yl)methanone

A solution of the product of Step H (405 mg, 0.83 mmol, 1 eq) and 5%Pd/C (100 mg) in ethanol (10 mL) at rt was placed under a balloon ofhydrogen gas overnight. The suspension was filtered through celite,washed with methanol, and the filtrates concentrated to give the productof Step I as a gum. Calculated for C18H31N3O4: 354.2 (M+1). found:354.2.

Step J((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone

A solution of the product of Step I (40 mg, 0.11 mmol, 1 eq), DIEA (0.06mL, 0.34 mmol, 3 eq) and 2-chloro-4-(trifluoromethyl)pyrimidine (0.04mL, 0.34 mmol, 3 eq) in a mixture of 10:1 dioxane/DMSO (1 mL) in a vialunder Ar was heated to 100° C. overnight. Water was added, the solutionwas extracted with DCM, the organics combined, dried over MgSO₄ andconcentrated. Purification by chromatography (4 g column) eluting with 5to 10% MeOH/DCM afforded the title compound of Example 23. ¹H NMR(CHLOROFORM-d) δ: 8.53 (d, J=4.6 Hz, 1H), 6.83 (d, J=4.9 Hz, 1H), 5.05(d, J=4.4 Hz, 1H), 4.10 (dd, J=12.3, 2.8 Hz, 1H), 3.51-4.00 (m, 13H),3.36-3.47 (m, 4H), 3.25-3.36 (m, 2H), 2.78 (dt, J=10.2, 3.8 Hz, 1H),2.34 (ddd, J=12.3, 6.7, 3.3 Hz, 1H), 2.20 (dt, J=13.0, 6.6 Hz, 2H), 2.01(dt, J=12.3, 8.3 Hz, 1H), 1.45-1.90 (m, 4H). Calculated forC23H32F3N5O4: 500.2 (M+1). found: 500.3.

Example 24 ((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(4-(6-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methanone

The title compound of Example 24 was made by taking the product ofExample 23 Step I and reacting with 2-chloro-6-(trifluoromethyl)pyridinefollowing the procedure described in Example 23, Step J. ¹H NMR(CHLOROFORM-d) δ: 7.63 (t, J=8.1 Hz, 1H), 7.01 (d, J=7.3 Hz, 1H), 6.81(d, J=8.6 Hz, 1H), 5.05 (d, J=4.4 Hz, 1H), 4.09 (dd, J=12.3, 3.1 Hz,1H), 3.96 (qd, J=7.8, 3.5 Hz, 2H), 3.47-3.88 (m, 10H), 3.37-3.47 (m,4H), 3.26-3.37 (m, 2H), 2.78 (dt, J=10.2, 3.6 Hz, 1H), 2.34 (ddd,J=12.2, 6.7, 3.4 Hz, 1H), 2.13-2.27 (m, 2H), 2.01 (dt, J=12.3, 8.5 Hz,1H), 1.60-1.88 (m, 4H), 1.54 (ddd, J=13.1, 10.9, 4.8 Hz, 1H). Calculatedfor C24H33F3N4O4: 499.3 (M+1). found: 499.4.

Example 25((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)((1S,4S)-5-(4-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methanone

The title compound of Example 25 was made by taking the product ofExample 23, Step E and reacting with(1S,4S)—N-Cbz-2,5-diaza-bicyclo[2.2.1]heptane following the proceduredescribed in Example 23, Step F, and then following the proceduresdescribed in Example 23, Steps G through J. ¹H NMR (CHLOROFORM-d) δ:8.50 (d, J=4.4 Hz, 1H), 6.83 (d, J=4.6 Hz, 1H), 4.70-5.20 (m, 3H), 4.07(t, J=10.1 Hz, 1H), 3.83-4.00 (m, 2H), 3.50-3.79 (m, 5H), 3.22-3.48 (m,7H), 2.79 (d, J=9.5 Hz, 1H), 1.82-2.30 (m, 7H), 1.40-1.82 (m, 4H).Calculated for C24H32F3N5O4: 512.2 (M+1). found: 512.3.

Example 26((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)((1S,4S)-5-(6-(trifluoromethyl)pyridin-2-yl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methanone

The title compound of Example 26 was made by taking the product ofExample 23, Step E and reacting with(1S,4S)—N-Cbz-2,5-diaza-bicyclo[2.2.1]heptane following the proceduredescribed in Example 23, Step F, then following the procedures describedin Example 23, Steps G through I, and then reacting that product with2-chloro-6-(trifluoromethyl)pyridine following the procedure describedin Example 23, Step J. ¹H NMR (CHLOROFORM-d) δ: 7.56 (t, J=7.9 Hz, 1H),6.94 (dd, J=7.0, 3.5 Hz, 1H), 6.40-6.57 (m, 1H), 4.65-5.18 (m, 3H), 4.05(d, J=12.5 Hz, 1H), 3.82-3.99 (m, 2H), 3.61-3.70 (m, 2H), 3.19-3.59 (m,10H), 2.53-2.85 (m, 1H), 1.84-2.29 (m, 7H), 1.44-1.78 (m, 4H).Calculated for C25H33F3N4O4: 511.3 (M+1). found: 511.2.

Example 27((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)((1S,4S)-5-(2-(trifluoromethyl)pyrimidin-4-yl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methanone

The title compound was made by taking the product of Example 23, Step Eand reacting with (1S,4S)—N-Cbz-2,5-diaza-bicyclo[2.2.1]heptanefollowing the procedure described in Example 23, Step F, then followingthe procedures described in Example 23, Steps G through I, and thenreacting that product with 4-chloro-2-(trifluoromethyl)pyrimidinefollowing the procedure described in Example 23, Step J. ¹H NMR(CHLOROFORM-d) δ: 8.27-8.43 (m, 1H), 6.24-6.60 (m, 1H), 5.25-5.48 (m,1H), 4.50-5.23 (m, 2H), 4.07 (d, J=12.2 Hz, 1H), 3.85-4.02 (m, 2H),3.58-3.73 (m, 3H), 3.23-3.56 (m, 9H), 2.55-2.91 (m, 1H), 1.86-2.35 (m,7H), 1.35-1.82 (m, 4H). Calculated for C24H32F3N5O4: 512.2 (M+1). found:512.2.

Example 28((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(4-(2-(trifluoromethyl)pyrimidin-4-yl)piperazin-1-yl)methanone

The title compound was made by taking the product of Example 23, Step Iand reacting with 4-chloro-2-(trifluoromethyl)pyrimidine following theprocedure described in Example 23, Step J. ¹H NMR (CHLOROFORM-d) δ: 8.37(d, J=6.4 Hz, 1H), 6.63 (d, J=6.1 Hz, 1H), 5.04 (d, J=4.6 Hz, 1H), 4.10(dd, J=12.2, 2.7 Hz, 1H), 3.90-4.02 (m, 2H), 3.47-3.90 (m, 10H),3.36-3.46 (m, 4H), 3.26-3.36 (m, 2H), 2.77 (dt, J=10.1, 3.7 Hz, 1H),2.32 (ddd, J=12.1, 6.7, 3.4 Hz, 1H), 2.19 (td, J=12.8, 6.2 Hz, 2H),1.95-2.08 (m, 1H), 1.59-1.86 (m, 4H), 1.54 (ddd, J=13.1, 11.1, 4.9 Hz,1H). Calculated for C23H32F3N5O4: 500.2 (M+1). found: 500.2.

Example 29((3aS,5S,6aR)-5-((3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethoxy)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound was prepared from reaction of the product of Example23, Step E and 3-(trifluoromethoxy)-5,6,7,8-tetrahydro-1,6-naphthyridinefollowing the procedure described in Example 1, Step C, and thenfollowing procedures described in Example 1, Steps G and H. ¹H NMR(CHLOROFORM-d) δ: 8.40 (br. s., 1H), 7.35 (br. s., 1H), 5.05 (t, J=4.3Hz, 1H), 4.74 (br. s., 2H), 3.78-4.16 (m, 5H), 3.48-3.74 (m, 2H),3.24-3.46 (m, 6H), 3.08 (br. s., 2H), 2.81 (t, J=9.3 Hz, 1H), 2.17-2.40(m, 3H), 1.91-2.12 (m, 3H), 1.53-1.88 (m, 3H). Calculated forC23H30F3N3O5: 486.2 (M+1). found: 486.1.

Example 30((3aS,5S,6aR)-5-((3S,4S)-3-methoxytetrahydro-2H-pyran-4-ylamino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanonesuccinate

Step A (1R,4S)-methyl 4-aminocyclopent-2-enecarboxylate hydrochloride

A solution of (1S,4R)-2-azabicyclo[2.2.1]hept-5-en-3-one (725 g, 6.64mol) in MeOH (2.2 L) was stirred in an ice bath to 0° C. Thionylchloride (290 mL, 3.99 mol) was added dropwise over a 2.25 h periodwhile keeping the temperature below 13° C. The reaction was stirred for2 h at 8° C. Isopropyl acetate (16.3 L) was added and the slurry stirredfor 1 h. The solid was filtered with a Buchner funnel, washed withisopropylacetate (˜1 L) and the solid was allowed to air-dry overnightto afford an off-white solid. ¹H NMR (400 MHz, DMSO-d₆) δ=8.44 (br. s.,3H), 5.99-6.16 (m, 1H), 5.90 (dt, J=2.4, 5.3 Hz, 1H), 4.17 (br. s., 1H),3.56-3.79 (m, 4H), 2.56 (m, 1H), 1.84-2.04 (m, 1H).

Step B (1R,4S)-methyl4-(tert-butoxycarbonylamino)cyclopent-2-enecarboxylate

A solution of the product of Step A (551 g, 3.10 mol), CH₂Cl₂ (15.5 L),and di-t-butyldicarbonate (684 g, 3.10 mol) was stirred to 2° C. with anice bath. Triethylamine (435 mL, 3.12 mol) was added over 1 h 5 min at arate not to exceed 3° C. The reaction was stirred for 2 h. The volatileswere evaporated, the crude product was suspended in a mixture of EtOAcand heptane, the solid was filtered through silica gel, and washed withadditional EtOAc in heptane. The organics were evaporated and affordedthe product of Step B as a brown solid. ¹H NMR (400 MHz,) δ=5.87 (d,J=6.4 Hz, 2H), 4.85-5.02 (m, 1H), 4.72-4.85 (m, 1H), 3.72 (s, 3H), 3.47(m, 1H), 2.51 (d, J=13.9 Hz, 1H), 1.88 (s, 1H), 1.44 (s, 10H).

Step C (1,1-dimethylethyl)(2-iodoethoxy)dimethylsilane

Iodoethanol (2.68 kg, 15.4 mol), CH₂Cl₂ (12 L) and imidizaole (1.556 kg,22.63 mol) were chilled in an ice bath. A solution oft-butyldimethylchlorosilane (2.536 kg, 16.32 mol) in CH₂Cl₂ (2.5 L) wasadded to the reaction over a 2 h period. The resulting white suspensionwas allowed to warm to rt over an 18 h. The reaction was worked up bywashing with water and brine). The organic layer was dried (MgSO₄) andevaporated under reduced pressure to provide the product of Step C as alight yellow oil. ¹H NMR (400 MHz, CDCl₃) δ=3.75 (t, J=7.0 Hz, 2H), 3.11(t, J=7.0 Hz, 2H), 0.77-0.89 (m, 10H), 0.00 (s, 6H).

Step D (1S,4S)-methyl4-(tert-butoxycarbonylamino)-1-(2-(tert-butyldimethylsilyloxy)ethyl)cyclopent-2-enecarboxylate

To a −70° C. solution of LiHMDS in THF (1M, 6.97 L, 6.97 mol) was addeda solution of the product of Step B (763.5 g, 3.16 mol) in THF (800 mL)over a 2 h period while keeping the temperature at or below −68° C. Theresulting solution was stirred for 45 min at −68° C. A solution of theproduct of Step C (1.267 kg, 4.426 mol) in THF (800 mL) was added over 1h 50 min period while maintaining a temperature of ˜−66° C. The reactionwas stirred at ˜−66° C. for 45 min. The reaction was warmed to −15° C.and worked up by addition to mixture of aqueous HCl and ice. The mixturewas extracted with toluene, the organic layer was washed with water,brine and dried over MgSO₄. The organic layer was concentrated andpurified on silica gel using a mixture of EtOAc in heptanes to providethe product of Step D as a clear oil. ¹H NMR (400 MHz, CHLOROFORM-d)d=5.69-5.86 (m, 2H), 4.79-4.93 (m, 1H), 4.68-4.80 (m, 1H), 3.67 (s, 3H),3.53-3.62 (m, 2 H), 2.16-2.30 (m, 1H), 2.04-2.16 (m, 2H), 1.70-1.81 (m,1H), 1.41 (s, 9H), 0.78-0.91 (m, 13H), 0.00 (s, 6H).

Step E tert-Butyl (5S,7S)-1-oxo-2-oxaspiro[4.4]non-8-en-7-ylcarbamate

To a solution of the product of Step D (1149 g, 2.875 mol) and THF (5.75L) was added TBAF (1M in THF, 2.875 L) over ˜1 h. The reaction wasstirred for 1 h at rt, and diluted with EtOAc. The organic layer waswashed with brine, diluted with heptanes and the organic layer wasfurther washed with water and brine. The organic layer was evaporated,the crystallized product was filtered, and washed with heptanes toprovide the product of Step E as a white solid. ¹H NMR (400 MHz,CHLOROFORM-d) d=5.86-5.98 (m, 3H), 5.67 (d, J=5.4 Hz, 3H), 5.03-5.20 (m,2H), 4.76-4.87 (m, 3H), 4.28 (t, J=7.0 Hz, 5H), 2.08-2.31 (m, 8H), 1.99(d, J=2.4 Hz, 3H), 1.34 (s, 25H).

Step F tert-butyl((1S,4S)-4-(2-hydroxyethyl)-4-(hydroxymethyl)cyclopent-2-en-1-yl)carbamate

To a solution of the product of Step E (255.9 g, 1.01 mol) and MeOH (2L) chilled to 2° C. was added NaBH₄ (75 g) over ˜2.5 h. The reaction wasquenched by addition of aqueous NH₄Cl, concentrated under reducedpressure and the mixture was diluted with water and EtOAc. The layerswere separated and the aqueous layer was extracted with additionalEtOAc. The combined organics were washed with brine, dried (MgSO₄) andevaporated to give the product of Step F as a thick oil. ¹H NMR (400MHz, CHLOROFORM-d) d=5.74 (d, J=2.0 Hz, 2H), 4.81-4.92 (m, 1H),4.67-4.79 (m, 1H), 3.71 (t, J=6.1 Hz, 2H), 3.50 (d, J=11.7 Hz, 2H),2.14-2.28 (m, 2H), 1.70 (td, J=1.6, 6.2 Hz, 4H), 1.52-1.60 (m, 1H), 1.44(s, 9H)

Step G tert-butyl (3aR,5S,6S,6aS)-6-bromo-3a-(hydroxymethyl)hexahydro-2H-cyclopenta[b]furan-5-ylcarbamate

To a chilled solution of the product of Step F (343.60 g) in EtOAc (4 L)was added N-bromosuccinimide (237.60 g) followed by stirring at rt for18 h. To the mixture was added water (5 mL) and the reaction heated to60° C. for 30 min. The reaction was filtered, the filtrate was washedwith aqueous sodium thiosulfate until the organic layer was negative forperoxides. The organic layer was washed with aqueous Na₂CO₃ (10%), dried(Na₂SO₄) and the reaction was concentrated under reduced pressure. Nearthe end of the concentration, heptane (1.2 L) was added, and the productwas collected by filtration to provide the product of Step G. ¹H NMR(400 MHz, CHLOROFORM-d) δ=4.78-4.91 (m, 1H), 4.41 (br. s., 1H), 4.31 (s,2H), 3.88-3.98 (m, 1H), 3.61-3.77 (m, 3H), 2.08-2.24 (m, 1H), 1.82 (m,2H), 1.60-1.70 (t, 1H), 1.45 (s, 9H)

Step H tert-Butyl(3aR,5S,6aR)-3a-(hydroxymethyl)hexahydro-2H-cyclopenta[b]furan-5-ylcarbamate

A solution of the product of Step G (83 g, 0.245 mol), 10% Pd on C (12.5g), triethylamine (69 mL, 0.49 mol, 2 eq.) in EtOAc (830 mL) was shakenon a PAAR hydrogenator at 40 psi for 3.5 h until the pressure remainedconstant. The reaction was filtered with celite, the filter cake waswashed with EtOAc, and the collected filtrate was washed with aqueousHCl (1N), brine, dried (Na₂SO₄) and concentrated under reduced pressureto give the product of Step H. ¹H NMR (400 MHz, CHLOROFORM-d)d=4.60-4.73 (m, 1H), 4.06-4.24 (m, 3H), 3.84-3.93 (m, 1H), 3.52-3.67 (m,4H), 2.12-2.21 (m, 1H), 1.98-2.04 (m, 1H), 1.90 (br. s., 3H), 1.70-1.78(m, 1H), 1.46-1.56 (m, 3H), 1.44 (s, 11H).

Step I(3aS,5S,6aR)-5-(tert-Butoxycarbonylamino)hexahydro-2H-cyclopenta[b]furan-3a-carboxylicacid

To an ice-cold solution of the product of Step H (426 g, 1.57 mol) andacetone (8.1 L) was added Jones reagent (710 mL) over 1 h 20 min. Theresulting suspension was stirred at rt for 1 h, after which isopropylalcohol (40 mL) was added, and the reaction stirred for 25 min at rt.Water was added, and the water/acetone was decanted off and evaporated.The insoluble material was dissolved separately in water and extractedwith CH₂Cl₂. The green water/acetone concentrate was extracted withCH₂Cl₂ and the combined organic extracts were washed with brine, dilutedwith water and EtOAc and the organic layer was dried with Na₂SO₄. Theorganic layer was filtered, concentrated and the product wascrystallized from MeCN, and the product of Step I was isolated byfiltration as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ=12.39-12.59 (m,1H), 6.85-7.02 (m, 1H), 4.28 (d, J=5.4 Hz, 1H), 3.89-3.97 (m, 1H),3.79-3.85 (m, 1H), 3.43-3.51 (m, 1H), 3.30-3.36 (m, 1H), 2.35-2.42 (m,1H), 1.90 (d, J=11.0 Hz, 3H), 1.59-1.70 (m, 1H), 1.42-1.50 (m, 1H), 1.37(s, 9H). Elemental anal calc for C13H21NO5: C, 57.55; H, 7.80; N, 5.16.Found: C, 57.34; H, 8.18; N, 5.08 mp: 147.4-149.1° C.

Step J tert-Butyl (3aS,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-ylcarbamate

To a solution of the product of Step I (596.8 g, 1.91 mol) in CH₂Cl₂ wasadded EDC (98.5% pure, 559 g, 2.87 mol) and HOBt (449 g, 3.26 mol) andthe suspension was stirred for 15 min at rt.3-(Trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-2HCl (790 g,2.87 mol) was added, followed by DIEA (1.7 L, 9.65 mol) by addition over45 min. The reaction was stirred at rt for 20 h. The reaction waspartitioned between saturated aqueous NaHCO₃ and CH₂Cl₂. The organiclayer was removed, the aqueous layer diluted with water and the aqueouslayer extracted with CH₂Cl₂. The combined organics were washed with ½saturated brine, dried over Na₂SO₄, and concentrated under reducedpressure. The resulting crude product was purified by chromatographyusing EtOAc in heptane to provide the product of Step J as a thickorange foam. ¹H NMR (400 MHz, CHLOROFORM-d) d=8.72 (s, 1H), 7.71 (s,1H), 5.00-5.08 (m, 1H), 4.77 (br. s., 2H), 4.61-4.68 (m, 1H), 4.21-4.34(m, 1H), 3.96-4.05 (m, 1H), 3.85-3.93 (m, 2H), 3.71 (s, 1H), 3.13 (br.s., 2H), 2.38-2.48 (m, 1H), 2.28-2.33 (m, 1H), 2.20-2.26 (m, 1H),2.09-2.17 (m, 1H), 1.75-1.85 (m, 1H), 1.61-1.70 (m, 1H), 1.40 (s, 9H).

Step K((3aS,5S,6aR)-5-Aminohexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanonedihydrochloride

A solution of the product of Step J (773 g, 1.61 mol) and HCl in MeOH(˜1.25 M, 14.25 L, 17.81 mol) was heated to 60° C. and after thevigorous bubbling ceased, the reaction was concentrated under reducedpressure. Isopropyl alcohol was added, the contents were evaporated tonear-dryness and heptane was added to the flask. The contents werefiltered, washed with some isopropyl alcohol/heptane (ad lib) and thesolid was dried in air, followed by drying in a vacuum oven to affordthe product of Step K as an ivory solid. A small sample of the productwas converted to the free-based using 1,2-dichloroethane/aqueous 3M NaOHfor further analysis by NMR and elemental analysis. Elemental analysiscalc for C₁₇H₂₀F₃N₃O₂×1.6 H₂O: C, 53.14; H, 6.09; F, 14.83; N, 10.93;H₂O=7.50. Found: C, 52.30; H, 5.78; F, 14.62; N, 10.51; KF=7.28. ¹H NMR(400 MHz, CHLOROFORM-d) δ=8.72 (s, 1H), 7.70 (br. s., 1H), 5.06 (d,J=4.9 Hz, 1H), 4.78 (s, 2H), 3.85-4.03 (m, 3H), 3.71-3.75 (m, 2H),3.59-3.70 (m, 2H), 3.08-3.19 (m, 2H), 2.24-2.38 (m, 2H), 2.19 (dd,J=5.7, 13.3 Hz, 1H), 2.08 (br. s., 1H), 1.62-1.78 (m, 1H), 1.46-1.57 (m,1H), 1.42 (br. s., 3H)

Step L((3aS,5S,6aR)-5-((3S,4S)-3-Methoxytetrahydro-2H-pyran-4-ylamino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

To a mixture of the product of Step K (the free base) (619.7 g, 1.74mol) in 1,2-dichloroethane/CH₂Cl₂ (˜10 L) was added acetic acid(glacial, 180 mL) and the mixture was cooled to 16° C. Solid Na(OAc)₃BH(463 g, 2.18 mol) was added and the suspension was stirred for 5-10 min.A solution of (R)-3-methoxydihydro-2H-pyran-4(3H)-one (prepared asdescribed in Intermediate 1, 213 g, 1.63 mol) in 1,2-dichloroethane(1.75 L) was added over 20 min and the resulting mixture was stirred atrt overnight. Additional acetic acid,(R)-3-methoxytetrahydro-4H-pyran-4-one (28 g) and Na(AcO)₃BH were addeduntil TLC showed the reaction was complete. The reaction was quenchedwith saturated aqueous NaHCO₃, the organic layer was separated and theaqueous layer was back-extracted with CH₂Cl₂. The combined organiclayers were washed with brine, dried (Na₂SO₄) and evaporated underreduced pressure. Purification was affected by chromatography with MeOH(7N NH₃) in CH₂Cl₂. The collected enriched isomer was further purifiedusing chiral chromatography on chiralpak AD column using a mixture ofheptanes/EtOH/isopropyl alcohol to provide the product of Step L.

¹H NMR (CHLOROFORM-d) δ: 8.72 (s, 1H), 7.70 (br. s., 1H), 5.05 (d, J=4.6Hz, 1H), 4.70-4.87 (m, 2H), 4.09 (dd, J=12.5, 2.7 Hz, 1H), 3.81-4.03 (m,4H), 3.62-3.71 (m, 1H), 3.50-3.62 (m, 1H), 3.35-3.46 (m, 4H), 3.24-3.35(m, 2H), 3.14 (t, J=4.9 Hz, 2H), 2.71-2.82 (m, 1H), 2.14-2.43 (m, 3H),1.99-2.13 (m, 1H), 1.46-1.86 (m, 5H). Calculated for C24H31F3N2O4: 470.2(M+1). found: 470.1.

Step M ((3aS,5S,6aR)-5-((3S,4S)-3-methoxytetrahydro-2H-pyran-4-ylamino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanonesuccinate

A solution of Example 30, Step L (608 g, 1.18 mol) in MeOH (6 L) waswarmed to 40° C. until dissolved. Succinic acid (141.9 g, 1.20 mol) wasadded and the suspension was warmed to 50° C. which caused everything todissolve. Darco G-60 charcoal (80 g) was added and the contents swirledfor 20 min. The mixture was filtered through Celite, washed with MeOH,and the solvent was evaporated under reduced pressure to provide thetitle compound (i.e., the succinate salt) as an amorphous foam. Theresulting foam was dissolved completely in MIBK (5 L, degassed) atreflux, the heating was stopped and the solution was allowed to cool.The solution was seeded at 104° C. with crystalline material, preparedas described in Example 52, and the solution was cooled to 38° C. over 4h. The suspension was chilled to 4° C., filtered, washed with ice-cold100 mL MIBK and the solid was allowed to dry under a positive nitrogenstream (protected from light) overnight. After some light milling, theproduct of Step M was collected as a white solid. ¹H NMR (400 MHz, MeOD)δ=8.72 (s, 1H), 8.04-8.12 (m, 1H), 4.97 (d, J=4.4 Hz, 1H), 4.94 (s, 3H),4.86 (s, 2H), 4.18-4.28 (m, 1H), 3.98 (d, J=11.7 Hz, 4H), 3.74-3.88 (m,1H), 3.62-3.73 (m, 1H), 3.28-3.58 (m, 8H), 3.12-3.23 (m, 2H), 2.58-2.68(m, 1H), 2.37-2.44 (m, 1H), 2.28-2.36 (m, 1H), 1.88 (m, 4H). ElementalAnalysis calc for C27H36F3N3O8×0.2 H₂O: C, 54.85; H, 6.21; F, 9.64; N,7.11; KF 0.61. Found: C, 55.17; H, 6.07; F, 9.99; N, 7.11; KF, 0.64.

Example 31((3aS,5S,6aR)-5-(4-phenylpiperidin-1-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step A1-((3aS,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)piperidin-4-one

To a suspension of sodium carbonate (2.2 g, 20.7 mmol, 5 eq) in methanol(70 mL) at 60° C. were added solutions of the product of Example 1, StepG (1.77 g, 4.14 mmol, 1 eq) in methanol (35 mL) 1,5-dichloropentan-3-one(0.74 g, 4.55 mmol, 1.1 eq) in methanol (35 mL) simultaneously over 1hour. After stirring 1 hr at 60° C., the suspension was cooled to rt,water was added, the methanol was concentrated, and the aqueousextracted with DCM, dried over MgSO₄ and concentrated. Purification bychromatography eluting with 2 to 6% MeOH/DCM afforded the title compoundof Step A. Calculated for C22H26F3N3O3: 438.2 (M+1). found: 438.2.

Step B 1-((3aS,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)-1,2,3,6-tetrahydropyridin-4-yltrifluoromethanesulfonate

To the product of Step A (1.06 g, 2.42 mmol, 1 eq) in THF (30 mL) at−78° C. under N₂ was added KHMDS (6.8 mL of a 0.5 M solution in toluene,3.39 mmol, 1.4 eq), the solution turned purple. After 15 minutes, asolution of N-phenyl-bis(trifluoromethanesulfonimide) (1.21 g, 3.39mmol, 1.4 eq) in THF (10 mL) was added and the yellow solution stirred 1hour at −78° C. Saturated NH₄Cl was added, the aqueous extracted withethyl acetate, dried over MgSO₄ and concentrated. Purification by columnchromatography (80 g) eluting with 3 to 6% MeOH/DCM afforded the titlecompound of Step B. Calculated for C23H25F6N3O5S: 570.1 (M+1). found:570.0.

Step C((3aS,5S,6aR)-5-(4-phenyl-5,6-dihydropyridin-1(2H)-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

A solution of the product of Step B (50 mg, 0.09 mmol, 1 eq),phenylboronic acid (22 mg, 0.18 mmol, 2 eq), (Ph₃P)₄Pd (10 mg, 0.009mmol, 0.1 eq) and 2 M Na₂CO₃ (0.1 mL) in dimethoxyethane (1 mL) under N₂was warmed to 80 C in a screw-top vial overnight. The solution wascooled to rt, and concentrated. Purification by column chromatography (4g) eluting with 50 to 100% ethyl acetate/heptane afforded the titlecompound of Step C. Calculated for C28H30F3N3O2: 498.2 (M+1). found:498.3.

Step D((3aS,5S,6aR)-5-(4-phenylpiperidin-1-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

A suspension of the product of Step D (24 mg, 0.046 mmol, 1 eq) and 5%Pd/C (20 mg) in ethanol (3 mL) was placed under a balloon of hydrogengas overnight. The solution was filtered through celite andconcentrated. Purification by column chromatography (4 g) eluting with 2to 6% MeOH/DCM afforded the title compound. ¹H NMR (CHLOROFORM-d) δ:8.71 (s, 1H), 7.70 (br. s., 1H), 7.27-7.33 (m, 2H), 7.15-7.24 (m, 3H),5.06 (d, J=4.5 Hz, 1H), 4.78 (br. s., 2H), 4.02 (td, J=8.1, 3.5 Hz, 1H),3.81-3.96 (m, 2H), 3.61-3.73 (m, 1H), 2.90-3.21 (m, 5H), 2.45-2.57 (m,1H), 2.22-2.32 (m, 2H), 2.08-2.15 (m, 3H), 1.99 (br. s., 2H), 1.65-1.89(m, 5H). Calculated for C28H32F3N3O2: 500.2 (M+1). found: 500.3.

Example 323-(1-((3aS,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)piperidin-4-yl)benzoicacid

The title compound of Example 32 was made by taking the product ofExample 31, Step B and reacting with 3-carboxyphenylboronic acidfollowing the procedure described in Example 31, Step C, then followingthe procedures described in Example 31, Step D. Calculated forC29H32F3N3O4: 544.2 (M+1). found: 544.0.

Example 33((3aS,5S,6aR)-5-(4-(3-methoxyphenyl)piperidin-1-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound of Example 33 was made by taking the product ofExample 31, Step B and reacting with 3-methoxyphenylboronic acidfollowing the procedure described in Example 31, Step C, then followingthe procedures described in Example 31, Step D. Calculated forC29H34F3N3O3: 530.3 (M+1). found: 530.3.

Example 344-(1-((3aS,5S,6aR)-3a-(3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine-6-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)piperidin-4-yl)benzoicacid

The title compound of Example 34 was made by taking the product ofExample 31, Step B and reacting with 4-benzyloxycarbonylphenylboronicacid following the procedure described in Example 31, Step C, thenfollowing the procedures described in Example 31, Step D. ¹H NMR(CHLOROFORM-d) δ: 8.68 (s, 1H), 7.79-7.88 (m, J=8.1 Hz, 2H), 7.60 (br.s., 1H), 7.09-7.21 (m, J=8.6 Hz, 2H), 5.01 (d, J=4.0 Hz, 1H), 4.67-4.78(m, 1H), 4.51-4.67 (m, 1H), 4.02 (br. s., 1H), 3.63-3.96 (m, 3H),3.34-3.54 (m, 3H), 3.23 (d, J=11.6 Hz, 1H), 3.09 (br. s., 1H), 2.56-2.70(m, 2H), 2.27-2.50 (m, 5H), 1.84-2.20 (m, 6H). Calculated forC29H32F3N3O4: 544.2 (M+1). found: 544.2.

Example 35((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(6-(trifluoromethyl)-2H-benzo[e][1,3]oxazin-3(4H)-yl)methanone

Step A tert-butyl((3aS,5S,6aR)-3a-((2-(tert-butoxy)-5-(trifluoromethyl)benzyl)carbamoyl)hexahydro-2H-cyclopenta[b]furan-5-yl)carbamate

The product of Step A was prepared from the reaction of the product ofExample 23, Step E and(2-(tert-butoxy)-5-(trifluoromethyl)phenyl)methanamine preparedaccording to procedures in ACS Med. Chem. Letters 2010, 1, 14 followingthe procedure from Example 1, Step C. Calculated for C25H35F3N2O5: 523.2(M+23). found: 523.2.

Step B (3aS,5S,6aR)-5-amino-N-(2-hydroxy-5-(trifluoromethyl)benzyl)hexahydro-2H-cyclopenta[b]furan-3a-carboxamidedihydrochloride

A solution of the product of Step A (11.07 g, 20.57 mmol, 1 eq) inmethanol (60 mL) and solution of HCl in methanol (82 mL of 1.25 Msolution, 103 mmol, 5 eq) was heated to 55° C. for 2.5 days. Thesolution was concentrated to give the product of step B. Calculated forC16H19F3N2O3: 345.1 (M+1). found: 345.3.

Step C(3aS,5S,6aR)-5-(1,3-dioxoisoindolin-2-yl)-N-(2-hydroxy-5-(trifluoromethyl)benzyl)hexahydro-2H-cyclopenta[b]furan-3a-carboxamide

A solution of the product of Step B (8.58 g, 18.74 mmol, 1 eq), phthalicanhydride (5.55 g, 37.5 mmol, 2 eq) and DIEA (11.3 mL, 55.6 mmol, 3.5eq) in chloroform (150 mL) was heated to 70° C. for 2 hours. Thesolution was cooled to rt and carbonyl diimidazole (2.24 g 13.82 mmol, 3eq) was added and the solution heated to 60° C. for 2 hours. Thesolution was cooled to rt, 1 N HCl was added, the aqueous extracted withDCM, the organics combined, dried over MgSO4 and concentrated.Purification by chromatography (200 g column) eluting with 30 to 60 to80% EA/heptane afforded the product of Step C. Calculated forC24H21F3N2O5: 474.1 (M+1). found: 475.1.

Step D2-((3aS,5S,6aR)-3a-(6-(trifluoromethyl)-3,4-dihydro-2H-benzo[e][1,3]oxazine-3-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)isoindoline-1,3-dione

A solution of the product of Step C (7.97 g, 15.5 mmol, 1 eq),paraformaldehyde (9.28 g, 310 mmol, 20 eq) and p-toluenesulfonic acidhydrate (2.94 g, 15.5 mmol, 1 eq) in toluene (300 mL) was heated to 130°C. for 18 hours in a flask equipped with a Dean-Stark trap. The solutionwas cooled to rt and concentrated. Purification by chromatography (200 gcolumn) eluting with 25 to 60 to 100% ethyl acetate/heptane afforded theproduct of Step D. Calculated for C25H21F3N2O5: 487.1 (M+1). found:487.2.

Step E((3aS,5S,6aR)-5-aminohexahydro-2H-cyclopenta[b]furan-3a-yl)(6-(trifluoromethyl)-2H-benzo[e][1,3]oxazin-3(4H)-yl)methanone

A solution of the product of Step D (5.39 g, 11.1 mmol, 1 eq) andhydrazine (7.1 mL, 222 mmol, 20 eq) in ethanol (60 mL) was stirred at rt18 hours. The white solid was filtered, washed with methanol and DCM,and the filtrates concentrated. Saturated NaHCO₃ was added, the aqueousextracted with DCM, the organics combined, dried over MgSO₄ andconcentrated to afford the product of Step E. Calculated forC17H19F3N2O3: 357.1 (M+1). found: 357.3.

Step F((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(6-(trifluoromethyl)-2H-benzo[e][1,3]oxazin-3(4H)-yl)methanone

The title compound was prepared from the reaction of the product of StepE and (R)-3-methoxydihydro-2H-pyran-4(3H)-one (Intermediate 1) followingthe procedure described in Example 1, Step H. ¹H NMR (CHLOROFORM-d) δ:7.42 (d, J=8.6 Hz, 1H), 7.36 (br. s., 1H), 6.95 (d, J=8.1 Hz, 1H), 5.43(br. s., 2H), 4.96-5.11 (m, 1H), 4.82 (br. s., 2H), 3.84-4.15 (m, 3H),3.68 (d, J=7.1 Hz, 1H), 3.55 (br. s., 1H), 3.32-3.45 (m, 4H), 3.20-3.32(m, 2H), 2.69-2.84 (m, 1H), 2.39 (br. s., 1H), 2.12-2.25 (m, 2H),1.99-2.12 (m, 1H), 1.78-1.99 (m, 1H), 1.46-1.78 (m, 4H). Calculated forC23H29F3N2O5: 471.2 (M+1). found: 471.2.

Example 36((3aS,5S,6aR)-5-(((3R,4R)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(6-(trifluoromethyl)-2H-benzo[e][1,3]oxazin-3(4H)-yl)methanone

The title compound was prepared from the reaction of the product of StepE and (S)-3-methoxydihydro-2H-pyran-4(3H)-one following the proceduredescribed in Example 1, Step H. ¹H NMR (CHLOROFORM-d) δ: 7.42 (d, J=8.6Hz, 1H), 7.36 (br. s., 1H), 6.96 (d, J=8.6 Hz, 1H), 5.35-5.49 (m, 2H),5.05 (d, J=5.1 Hz, 1H), 4.74-4.90 (m, 2H), 4.05 (dd, J=12.6, 3.5 Hz,1H), 3.94-4.01 (m, 1H), 3.91 (dt, J=11.4, 3.7 Hz, 1H), 3.61-3.71 (m,1H), 3.48-3.60 (m, 1H), 3.24-3.41 (m, 6H), 2.78 (dd, J=6.3, 3.8 Hz, 1H),2.39 (br. s., 1H), 2.19 (td, J=12.1, 6.1 Hz, 2H), 2.05 (dt, J=12.3, 8.3Hz, 1H), 1.89 (br. s., 1H), 1.43-1.75 (m, 4H). Calculated forC23H29F3N2O5: 471.2 (M+1). found: 471.2.

Example 373-(1-((3aS,5S,6aR)-3a-(6-(trifluoromethyl)-3,4-dihydro-2H-benzo[e][1,3]oxazine-3-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)piperidin-4-yl)benzoicacid

The title compound of Example 37 was made by taking the product ofExample 35, Step E following the procedures described in Example 31,Steps A and B, then reacting that product with3-benzyloxycarbonylphenylboronic acid following the procedure describedin Example 31, Step C, then following the procedure described in Example31, Step D. ¹H NMR (CHLOROFORM-d) δ: 8.04 (s, 1H), 7.78 (d, J=6.8 Hz,1H), 7.32-7.43 (m, 2H), 7.19-7.32 (m, 2H), 6.93 (d, J=8.3 Hz, 1H),5.16-5.66 (m, 2H), 5.06 (d, J=4.2 Hz, 1H), 4.96 (d, J=16.9 Hz, 1H), 4.72(d, J=16.9 Hz, 1H), 3.96-4.11 (m, 1H), 3.55-3.75 (m, 2H), 3.47 (s, 1H),3.29-3.45 (m, 2H), 2.59-2.80 (m, 2H), 2.35-2.59 (m, 5H), 2.30 (dd,J=12.8, 5.7 Hz, 1H), 2.13-2.24 (m, 1H), 1.99-2.13 (m, 2H), 1.92 (d,J=11.7 Hz, 1H), 1.82 (d, J=13.4 Hz, 1H). Calculated for C29H31F3N2O5:545.2 (M+1). found: 545.2.

Example 38((3aS,5S,6aR)-5-(4-phenylpiperidin-1-yl)hexahydro-2H-cyclopenta[b]furan-3a-yl)(6-(trifluoromethyl)-2H-benzo[e][1,3]oxazin-3(4H)-yl)methanone

The title compound of Example 38 was made by taking the product ofExample 35, Step E following the procedures described in Example 31,Steps A through D. ¹H NMR (CHLOROFORM-d) δ: 7.42 (d, J=8.6 Hz, 1H), 7.37(br. s., 1H), 7.29 (t, J=7.5 Hz, 2H), 7.15-7.24 (m, 3H), 6.96 (d, J=8.6Hz, 1H), 5.42 (br. s., 2H), 5.03 (d, J=4.6 Hz, 1H), 4.69-4.95 (m, 2H),3.95-4.07 (m, 1H), 3.60-3.73 (m, 1H), 3.04 (br. s., 3H), 2.32-2.56 (m,2H), 2.19-2.31 (m, 2H), 1.89-2.17 (m, 4H), 1.53-1.89 (m, 6H). Calculatedfor C28H31F3N2O3: 501.2 (M+1). found: 501.2.

Example 39N,N-dimethyl-3-(1-((3aS,5S,6aR)-3a-(6-(trifluoromethyl)-3,4-dihydro-2H-benzo[e][1,3]oxazine-3-carbonyl)hexahydro-2H-cyclopenta[b]furan-5-yl)piperidin-4-yl)benzamide

The title compound of Example 39 was made by taking the product ofExample 35, Step E following the procedures described in Example 31,Steps A and B, then reacting that product withN,N-dimethylbenzamide-3-boronic acid following the procedure describedin Example 31, Step C, then following the procedure described in Example31, Step D. Calculated for C31H36F3N3O4: 572.3 (M+1). found: 572.3.

Example 40((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)(methyl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound of Example 40 was made by taking the product ofExample 30 and reacting with formaldehyde following the proceduresdescribed in Example 1, Step H. ¹H NMR (CHLOROFORM-d) δ: 8.72 (br. s.,1H), 7.70 (br. s., 1H), 5.03 (d, J=4.6 Hz, 1H), 4.67-4.92 (m, 2H), 4.15(d, J=12.7 Hz, 1H), 3.97-4.10 (m, 2H), 3.92 (br. s., 2H), 3.55-3.71 (m,2H), 3.33-3.53 (m, 5H), 3.22 (d, J=12.7 Hz, 1H), 3.14 (br. s., 2H), 2.64(d, J=11.7 Hz, 1H), 2.22-2.50 (m, 4H), 1.79-2.22 (m, 6H), 1.69 (td,J=12.5, 4.9 Hz, 1H), 1.51 (d, J=12.0 Hz, 1H). Calculated forC24H32F3N3O4: 484.2 (M+1). found: 484.2.

Example 41((3aS,5S,6aR)-5-((2-methoxyethyl)((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound of Example 41 was made by taking the product ofExample 30 and reacting with methoxyacetaldehyde following theprocedures described in Example 1, Step H. Calculated for C26H36F3N3O5:528.3 (M+1). found: 528.3.

Example 42((3aS,5S,6aR)-5-(((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)(2,5,8,11-tetraoxatetradecan-14-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound of Example 42 was made by taking the product ofExample 30 and reacting with 4,7,10,13-tetraoxatetradecanal followingthe procedures described in Example 1, Step H. Calculated forC33H50F3N308: 674.4 (M+1). found: 674.4.

Example 43((3aS,5S,6aR)-5-((2-hydroxyethyl)((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

Step A((3aS,5S,6aR)-5-((2-((tert-butyldimethylsilyl)oxy)ethyl)((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound of Step A was made by taking the product of Example30 and reacting with t-butyldimethylsiloxyacetaldehyde following theprocedures described in Example 1, Step H. Calculated forC31H48F3N3O5Si: 628.3 (M+1). found: 628.2.

Step B ((3aS,5S,6aR)-5-((2-hydroxyethyl)((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

A solution of the product of Step A (210 mg, 0.33 mmol, 1 eq) in 1 N HCl(0.25 mL) and dioxane (5 mL) was heated to 90° C. for 18 hrs, thencooled to rt. Saturated NaHCO₃ was added, the aqueous extracted withCH₂Cl₂, dried over MgSO₄ and concentrated. Purification bychromatography (12 g) eluting with 3 to 8% MeOH/DCM afforded the titleof compound of Example 43 (67 mg, 38%). ¹H NMR (CHLOROFORM-d) δ: 8.72(br. s., 1H), 7.71 (br. s., 1H), 4.96 (d, J=4.4 Hz, 1H), 4.80 (br. s.,2H), 4.15 (d, J=12.7 Hz, 1H), 3.77-4.10 (m, 5H), 3.56-3.70 (m, 1H),3.30-3.52 (m, 7H), 3.23 (d, J=12.7 Hz, 1H), 3.14 (br. s., 2H), 2.66-2.90(m, 3H), 2.39 (br. s., 1H), 2.13-2.27 (m, 1H), 2.05 (dd, J=12.7, 5.6 Hz,3H), 1.82 (br. s., 1H), 1.68 (td, J=12.6, 4.9 Hz, 1H), 1.42 (br. s.,1H). Calculated for C25H34F3N3O5: 514.3 (M+1). found: 514.3.

Example 44((3aS,5S,6aR)-5-((3-hydroxypropyl)((3S,4S)-3-methoxytetrahydro-2H-pyran-4-yl)amino)hexahydro-2H-cyclopenta[b]furan-3a-yl)(3-(trifluoromethyl)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)methanone

The title compound of Example 44 was made by taking the product ofExample 30 and reacting with 3-((tert-butyldimethylsilyl)oxy)propanalfollowing the procedures described in Example 43 Steps A and B. ¹H NMR(CHLOROFORM-d) δ: 8.72 (s, 1H), 7.72 (s, 1H), 4.97 (d, J=4.5 Hz, 1H),4.69-4.91 (m, 2H), 4.15 (d, J=12.6 Hz, 1H), 3.83-4.10 (m, 5H), 3.58-3.82(m, 3H), 3.33-3.47 (m, 5H), 3.25 (d, J=12.6 Hz, 1H), 3.14 (br. s., 2H),2.70-2.93 (m, 3H), 2.36 (br. s., 1H), 2.15-2.30 (m, 1H), 1.97-2.15 (m,3H), 1.78-1.96 (m, 1H), 1.35-1.77 (m, 4H). Calculated for C26H36F3N3O5:528.3 (M+1). found: 528.3.

Example 45 In Vitro Biological Data

Compounds of the invention were subjected to various representativebiological tests. The results of these tests are intended to illustratethe invention in a non-limiting fashion. MCP-1 Receptor Binding Assay inTHP-1 Cells

Human monocytic cell line THP-1 cells were obtained from American TypeCulture Collection (Manassas, Va., USA). The THP-1 cells were grown inRPMI-1640 (RPMI: Roswell Park Memorial Institute Medium-cell culturegrowth media) supplemented with 10% fetal bovine serum in a humidified5% CO₂ atmosphere at 37° C. The cell density was maintained between0.5×10⁶ cells/mL.

THP-1 (cells were incubated with 0.5 nM ¹²⁵I labeled MCP-1 (Perkin-ElmerLife Sciences, Inc. Boston, Mass.) in the presence of varyingconcentrations of either unlabeled MCP-1 (R & D Systems, Minneapolis,Minn.) or test compound for 2 hours at 30° C. in a 96 well plate. Cellswere then harvested onto a filter plate, dried, and 20 μL of Microscint20 was added to each well. Plates were counted in a TopCount NXT,Microplate Scintillation & Luminescence Counter (Perkin-Elmer LifeSciences, Inc. Boston, Mass.). Blank values (buffer only) weresubtracted from all values and drug treated values were compared tovehicle treated values. 1 μM cold MCP-1 was used for nonspecificbinding.

Table 1 lists IC₅₀ values for inhibition of MCP-1 binding to CCR2obtained for test compounds of the invention. Where an IC₅₀ value wasnot obtained for a particular compound, the percent inhibition isprovided at a test concentration of 25 μM.

TABLE 1 Inhibition of MCP-1 Binding IC₅₀ Example CCR2 Binding (nM) 1 162 7 3 11 4 93 5 170 6 62 7 2 8 3 9 9 10 16 11 4 12 24 13 20 16 14 21 120

The compounds of Examples 15, 29-31, 33, 35-37, 39, and 40 are believedto have CCR2 binding of less than about 50 nM, those of Examples 14, 20,22, and 25 are believed to have CCR2 binding of about 50-100 nM, thoseof Examples 17, 26, 38, and 43-44 are believed to have CCR2 binding ofabout 100-200 nM, and those of Examples 18, 19, 23, 24, 27, 28, 32, 34,41 and 42 are believed to have CCR2 binding of greater than about 200nM.

Example 46 Animals

Mouse CCR2 knock-out/human CCR2 knock-in mice are generated usingtargeted 129Sv/Evbrd embryonic stem cell clones injected into C57BL/6mice. Expression of the hCCR2 transcript is confirmed by quantitativereverse transcription-polymerase chain reaction performed on spleen andblood total RNA from homozygous hCCR2 knock-in mice. Backcrossing intoC57BL/6 genetic background continued to the eighth generation.Transgenic mice are housed in a specific-pathogen-free,temperature-controlled facility that maintained a 12-hour light/12-hourdark cycle. Mice have free access to water and food. Experimentalprocedures are carried out in accordance with institutional standardsfor animal care and are approved by the institute's animal care and usecommittee.

Example 47 Murine In Vivo Cell Migration Assay

Animals are orally dosed with vehicle or CCR2 antagonists at 3, 10 and30 mg/kg bid. Animals undergo anesthesia and laparotomy. A distal loopof small bowel (5 cm in length) is gently eventrated onto moist sterilegauze. Synthetic human MCP-1 (1 mg/100 ml sterile PBS) or PBS alone isadministered drop-wise onto the serosa of the eventrated loop. A sutureknot is placed into the mesentery to mark the terminus of the treatedarea. Twenty-four hours later, the animal is sacrificed and the segmentof bowel plus the adjacent region is removed. The tissue is opened alongthe mesenteric border, pinned flat and the mucosa removed. The remainingmuscle layer is fixed briefly in 100% EtOH and then stained usingHanker-Yates reagent to detect myeloperoxidase-containing immune cells.At 10 mpk, P.O. bid, a compound is deemed efficacious if the inhibitionof cell migration reaches 30% compared with vehicle-treated animals.

Example 48 Thioglycollate-Induced Peritonitis in Mice

Animals were orally dosed with vehicle or the compound of Example 30 at0, 1, 3, and 10 mg/kg bid). One hour later, the animals wereintraperiponeally injected with sterile thioglycollate (25 mL/kg, ip,Sigma) for induction of peritonitis. Animals were orally treated twicedaily with vehicle or Example 30. At the 72-hour time point, perinotealcavities were lavaged with 10 mL of sterile saline. Total cell counts inthe peritoneal lavage fluid were performed using a microscope and celldifferentiation is performed using cytospin analysis after Giemsastaining (Hema Tek 2000). Percent inhibition of thethioglycollate-induced peritonitis was calculated by comparing thechange in number of leukocytes of CCR2 antagonist treated mice to thevehicle-treated mice. When the compound of Example 30 was administeredat 1, 3 and 10 mg/kg p.o bid, the thioglycollate induced cellularinfiltrate in hCCR2KI mice at 72 hr was inhibited by 51%, 67% and 95%,respectively. The effect of Example 30 was demonstrated to bedose-dependent with an ED₅₀ of 1 mg/kg p.o. bid, and a cmax EC₅₀ of 97nM in plasma (0.5 hour post the last dose).

Example 49 MCP-1-Induced Monocyte Recruitment to Airway of Mice

Animals are orally treated with vehicle or CCR2 antagonists at 3, 10,and 30 mg/kg po bid). One hour later, the animals are intranasally dosedwith 4 μg of MCP-1 in sterile saline. The animals are orally treatedtwice daily with vehicle or CCR2 antagonists. After 48 h, mice areeuthanized by intraperitoneal injection of anesthesia solution(Sleepaway-Sodium pentobarbital). Whole bronchoalveolar lavage (BAL) isperformed using 1.4 ml of ice-cold PBS containing 3 mM EDTA. Total cellcounts in the BAL lavage fluid are performed using a microscope and celldifferentiation is performed using cytospin analysis after Giemsastaining (Hema Tek 2000). Percent inhibition is calculated by comparingthe change in number of total leukocyte counts (includingmonocytes/macrophages and lymphocytes) of compound-treated mice to thevehicle-treated mice. Compounds are deemed efficacious if percentinhibition reaches 30%.

Example 50 High-Fat Diet Induced Obesity and Insulin Resistance in Mice

Obesity was induced by a high-fat diet that derived approximately 60%calories from fat (D-12492; Research Diets Inc.) in animals for 10-12weeks at age of 7 weeks. Prior to age 7 weeks, animals were fed astandard pellet diet, in which 5% of calories were provided as fat.Obese animals were randomized by body weight. The obese animals wereorally treated with vehicle or the compound of Example 30 at 1, 3, and10 mg/kg, po bid. Body weight and food intake and fasting blood glucoselevels were monitored. Body mass was determined by a NMR analyzer(Bruker MiniSpec). Insulin tolerance test was carried out in animalsthat fasted for 3 hours. After an intraperitoneal bolus injection ofrecombinant human insulin (0.5 U/kg), blood glucose concentrations weremeasured using a Glucometer before and 15, 30, 45, 60, 90 and 120minutes after injection. Glucose tolerance tests were performed after anovernight (17-hour) fast. Blood glucose concentrations were measuredbefore and after 15, 30, 60, 90, 120 minutes after an oral dose ofglucose dissolved in water (2.5 g/kg). Energy expenditure analysis wasmonitored by a complete laboratory animal monitor system. After 50 daystreatment with vehicle or CCR2 antagonists, the animals were sacrificedby CO₂ asphyxiation. Percent of weight loss was calculated by comparingthe body weight changes of the compound-treated mice with thevehicle-treated mice. After 32-days treatment, the compound of Example30 reduced the high fat-diet induced body weight by 4.94% (p>0.05),10.94% (p<0.01) and 15.7% (p<0.01) when administered at 1, 3 and 10mg/kg p.o. bid, respectively.

Example 51 Mouse Model of Allergic Asthma

Animals are sensitized by intraperitoneal injection of 10 μg chicken eggalbumin (OVA) absorbed to 1 mg Imject® in 100 μL phosphate-bufferedsaline (PBS) on days 0 and 5. Control animals received PBS ip.OVA-immunized animals are challenged by inhalation of 0.5% OVA aerosolfor 10 minutes by an ultrasonic nebulizer on days 12, 16 and 20. Controlanimals are challenged with PBS in similar fashion. The OVA-sensitizedanimals receive vehicle (0.5% Methocel) or CCR2 antagonists orally at 3,10, 30 mg/kg twice daily from days 9-20 and once daily on Day 21, 2hours before sacrifice. Dexamethason (5 mg/kg) and Montelukast (1 mg/kg)are given orally once a day. On day 21, 2 hours post the last dose ofCCR2 compounds, bronchial reactivity to aerosolized methacholine ismeasured using a Buxco whole body plethysmograpgh. On day 21, theanimals are sacrificed. Bronchoalveolar lavage fluid is collected (1 mL)and total cells counted. The numbers of eosinophils, lymphocytes,monocytes and neutrophils are determined using cytospin analysis afterGiemsa staining (Hema Tek 2000). Percent inhibition of total BALleukocyte count (and eosinophil count) is calculated by comparing thecompound-treated mice with vehicle-treated mice. Compounds are deemedefficacious if the inhibition reaches 30%.

Example 52 Preparation of Crystalline Succinate Salt of Compound ofFormula (I-S)

The crystalline succinate salt of the compound of formula (I) wasprepared by heating amorphous succinate salt (the amorphous succinatesalt is the foam described in Example 30, Step M) of the compound offormula (I) in an open DSC aluminum pan to about 140° C. with a heatingrate of 10° C./min, then cooling to about 30° C. with a cooling rate of10° C./min.

FIG. 5 illustrates a DSC thermogram measured during the experimentdescribed above. The DSC thermogram shows a first endothermic event atabout 50° C. (theorized to be the result of desolvation of the amorphousform); an exothermic event with a maximum at about 138° C., indicativeof crystallization; and a subsequent endothermic event at 155° C.,indicative of the melting of the crystalline solid.

FIG. 5 further includes a TGA thermogram for the amorphous succinatesalt of the compound of formula (I-S) used in the example describedabove, which shows about 4.8% weight loss between room temperature andabout 80° C.; and decomposition starting at about 172° C.

Example 53 Preparation of Crystalline Succinate Salt of Compound ofFormula (I-S)

The following general procedure was applied in a screening study foridentifying solvents suitable for crystallization of the crystallinesuccinate salt of the compound of formula (I-S). Crystalline succinatesalt of the compound of formula (I-S) was prepared from the amorphoussuccinate salt (the amorphous succinate salt is the foam described inExample 30, Step M) of the compound of formula (I-S), crystallizing frommethyl isobutyl ketone. (Note: Water, methanol ethanol, acetone,acetonitrile, isopropyl acetate, nitromethane, tetrahydrofuran, methylethyl ketone, dichloromethane, and toluene did not inducecrystallization.)

Amorphous succinate salt of the compound of formula (I-S) (5-10 mg) wassuspended in 1-2 mL of methyl-isobutyl ketone (MIBK). The resultingsuspension was heated in an oil bath and at reflux conditions, thesuspension formed a clear solution, which upon cooling to roomtemperature under ambient conditions yielded crystalline solids.

The solids isolated from MIBK were allowed to dry under ambientcondition and then analyzed by X-ray. The pXRD pattern of the solidisolated from MIBK was similar to the pXRD pattern of the heat treatedsample (prepared as in Example 52 above), indicating the samecrystalline form was produced in both cases.

Example 54 Oral Formulation—Prophetic Example

As a specific embodiment of an oral composition, 100 mg of the compoundprepared as in Example 53 is formulated with sufficient finely dividedlactose to provide a total amount of 580 to 590 mg to fill a size 0 hardgel capsule.

While the foregoing specification teaches the principles of the presentinvention, with examples provided for the purpose of illustration, itwill be understood that the practice of the invention encompasses all ofthe usual variations, adaptations and/or modifications as come withinthe scope of the following claims and their equivalents.

We claim:
 1. A compound of Formula (I):

wherein: A is O, or S; R⁰ is H, or C₍₁₋₄₎alkyl; wherein said C₍₁₋₄₎alkylis optionally substituted with OH, C₍₁₋₄₎alkyl-(OCH₂CH₂)_(n)—OCH₃, OCH₃,CO₂H, C(O)NH₂, SO₂NH₂, or CO₂C₍₁₋₄₎alkyl; n is 1, 2, or 3; R¹ iscyclohexyl, or tetrahydropyranyl; wherein said cyclohexyl ortetrahydropyranyl may be optionally substituted with one substituentselected from the group consisting of: OCH₃, OH, CH₂CH₃, —CN, NH₂,NH(CH₃), N(CH₃)₂, or OCF₃; alternatively, R⁰ and R¹ are taken togetherwith their attached nitrogen to form a ring selected from the groupconsisting of

R^(a) is phenyl; wherein the phenyl is optionally substituted withC(O)NH₂, C(O)NHC₍₁₋₄₎alkyl, SO₂NH₂, C(O)N(C₍₁₋₄₎alkyl)₂, OCH₃, CO₂CH₃,or CO₂H; R^(b) is C₍₁₋₄₎alkyl, or OC₍₁₋₄₎alkyl; R² is selected from thegroup consisting of H, C₍₁₋₄₎alkyl, cyclopropyl, cyclohexyl, phenyl,pyridyl, pyrimidyl, pyrazyl, pyrazolyl, imidazolyl, isoxazolyl,thiazolyl, furyl, and thiophenyl; wherein said phenyl, pyridyl,pyrimidyl, pyrazyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl,thiazolyl, furyl, or thiophenyl is optionally substituted with onesubstituent selected from the group consisting of NH₂, NHC₍₁₋₃₎alkyl,N(C₍₁₋₃₎alkyl)₂, C₍₁₋₃₎alkyl, —CN, —CH═CH₂, —CONH₂, —CO₂H, —NO₂,—CONHC₍₁₋₄₎alkyl, CON(C₍₁₋₄₎alkyl)₂, C₍₁₋₄₎alkylCONH₂, —NHCOC₍₁₋₄₎alkyl,—CO₂C₍₁₋₄₎alkyl, CF₃, SO₂C₍₁₋₄₎alkyl, —SO₂NH₂, —SO₂NH(C₍₁₋₄₎alkyl), and—SO₂N(C₍₁₋₄₎alkyl)₂; R³ is H, or CH₃; alternatively, R³ and R² are takentogether with their attached carbon to form

R⁴ is

R⁵ is H, or CH₃; alternatively, R⁴ and R⁵ are taken together with theirattached nitrogen to form a ring selected from the group consisting of:

R⁶ is CF₃, or OCF₃; R⁷ is a CF₃ substituted heteroaryl, provided that R⁷is not

R_(x) is CF₃, F, Cl, CN, or OCH₃; R_(y) is H, F, Cl, or CF₃; R_(z) is H,or F; or a pharmaceutically acceptable salt thereof.
 2. A compound ofclaim 1, wherein: A is O, or S; R⁰ is H, or C₍₁₋₄₎alkyl, wherein saidC₍₁₋₄₎alkyl is optionally substituted with OH,C₍₁₋₄₎alkyl(OCH₂CH₂)_(n)OCH₃, or OCH₃; n is 1, 2, or 3; R¹ iscyclohexyl, 1-methoxy cyclohex-2-yl, tetrahydropyran-4-yl, or 3-methoxytetrahydropyran-4-yl; alternatively, R⁰ and R¹ are taken together withtheir attached nitrogen to form a ring selected from the groupconsisting of

R^(a) is phenyl; wherein said phenyl is optionally substituted withC(O)NH₂, C(O)NHCH₃, SO₂NH₂, C(O)N(CH₃)₂, OCH₃, CO₂CH₃, or CO₂H; R² is H,C₍₁₋₄₎alkyl, cyclopropyl, cyclohexyl, thiazol-2-yl,1-methyl-imidazol-2-yl, 1-methyl-pyrazol-5-yl, or phenyl; R⁴ and R⁵ aretaken together with their attached nitrogen to form a ring selected fromthe group consisting of:

R⁶ is CF₃, or OCF₃; R⁷ is

or a pharmaceutically acceptable salt thereof.
 3. A compound of claim 2,wherein: A is O, or S; R⁰ is H, CH₃, CH₂CH₂CH₂OH, CH₂CH₂OH,CH₂CH₂CH₂—(OCH₂CH₂)₃—OCH₃, or CH₂CH₂OCH₃; R¹ is tetrahydropyran-4-yl, or3-methoxy tetrahydropyran-4-yl; alternatively, R⁰ and R¹ are takentogether with their attached nitrogen to form a ring selected from thegroup consisting of

R^(a) is phenyl; wherein said phenyl is optionally substituted withC(O)N(CH₃)₂, OCH₃, or CO₂H; R⁴ and R⁵ are taken together with theirattached nitrogen to form a ring selected from the group consisting of:

R⁶ is CF₃, or OCF₃; R⁷ is

or a pharmaceutically acceptable salt thereof.
 4. A compound of claim 3,wherein: A is O; or a pharmaceutically acceptable salt thereof.
 5. Acompound of claim 1 selected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 6. A compound which is

and pharmaceutically acceptable salts thereof.
 7. A succinate salt of acompound of formula (I-S)


8. A salt as in claim 7, wherein the salt is crystalline.
 9. A salt asin claim 7, wherein the salt is a crystalline hydrate; and wherein thehydrate contains about 0.6 moles water per mole of the compound offormula (I-S).
 10. A salt as in claim 7, wherein the salt is acrystalline hydrate; and wherein the salt is hygroscopic.
 11. A salt asin claim 7, wherein the salt is crystalline and exhibits a peaktemperature of melting, as measured by DSC, of about 158° C.
 12. Acrystalline succinate salt of a compound of formula (I-S)

comprising powder X-ray diffraction peaks of 11.27, 13.87, 19.22 and22.01° 2θ.
 13. A pharmaceutical composition, comprising a compound ofclaim 1 and a pharmaceutically acceptable carrier.
 14. A pharmaceuticalcomposition made by mixing a compound of claim 1 and a pharmaceuticallyacceptable carrier.
 15. A process for making a pharmaceuticalcomposition comprising mixing a compound of claim 1 and apharmaceutically acceptable carrier.
 16. A method for preventing,treating or ameliorating a CCR2 mediated syndrome, disorder or diseasecomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of claim
 1. 17. A method for preventing,treating or ameliorating a CCR2 mediated inflammatory syndrome, disorderor disease wherein the syndrome, disorder or disease is associated withelevated MCP-1 expression or MCP-1 overexpression, or is an inflammatorycondition that accompanies syndromes, disorders or diseases associatedwith elevated MCP-1 expression or MCP-1 overexpression comprisingadministering to a subject in need thereof an effective amount of acompound of claim
 1. 18. A method of preventing, treating orameliorating a syndrome, disorder or disease, wherein said syndrome,disorder or disease is selected from the group consisting of: chronicobstructive pulmonary disorder (COPD), ophthalmic disorders, uveitis,atherosclerosis, rheumatoid arthritis, psoriasis, psoriatic arthritis,atopic dermatitis, multiple sclerosis, Crohn's Disease, ulcerativecolitis, nephritis, organ allograft rejection, fibroid lung, renalinsufficiency, type-I diabetes, type II diabetes, diabeticcomplications, diabetic nephropathy, diabetic retinopathy, diabeticretinitis, diabetic microangiopathy, overweight, obesity,obesity-associated insulin resistance, metabolic syndrome, tuberculosis,sarcoidosis, invasive staphyloccocia, inflammation after cataractsurgery, allergic rhinitis, allergic conjunctivitis, chronic urticaria,asthma, allergic asthma, periodontal diseases, periodonitis, gingivitis,gum disease, diastolic cardiomyopathies, cardiac infarction,myocarditis, chronic heart failure, angiostenosis, restenosis,reperfusion disorders, aortic abdominal aneurism, glomerulonephritis,solid tumors and cancers, chronic lymphocytic leukemia, chronicmyelocytic leukemia, multiple myeloma, malignant myeloma, Hodgkin'sdisease, and carcinomas of the bladder, breast, cervix, colon, lung,prostate, or stomach, and chronic neuroinflammatory disorders including,but not limited to, Alzheimer's disease, ischemic stroke, spinal cordinjury, nerve crush injury and traumatic brain injury comprisingadministering to a subject in need thereof an effective amount of acompound of claim
 1. 19. A method of preventing, treating orameliorating a syndrome, disorder or disease, wherein said syndrome,disorder or disease is selected from the group consisting of: type Idiabetes, type II diabetes, diabetic complications, diabeticnephropathy, diabetic retinopathy, diabetic retinitis, diabeticmicroangiopathy, obesity, obesity-associated insulin resistance,metabolic syndrome, asthma, and allergic asthma, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of claim
 1. 20. A method of treating a disorderselected from the group consisting of type II diabetes, obesity andasthma comprising administering to a subject in need thereof atherapeutically effective amount of a compound of claim
 1. 21. Acompound of formula (XXI)


22. A process for making (S)-4,4-dimethoxytetrahydro-2H-pyran-3-olcomprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and R-(+)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 60% enantiomeric excess.
 23. The process of claim 22 wherein(S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol is formed in at least 90%enantiomeric excess.
 24. The process of claim 22 wherein the boranereducing complex is selected from borane-dimethylsulfide complex orborane-N,N-diethylaniline complex.
 25. The process of claim 24 wherein aTHF solution of the borane-reducing complex andR-(+)-2-methyl-CBS-oxazaborolidine is added to a solution of the4-dimethoxydihydro-2H-pyran-3(4H)-one in THF.
 26. The process of claim22 wherein the reaction is carried out in an inert environment.
 27. Theprocess of claim 26 wherein the reaction is run under nitrogen gas. 28.The reaction of claim 22 further comprising reacting the(S)-4,4-dimethoxytetrahydro-2H-pyran-3-ol with dimethyl sulfate toprovide (R)-3,4,4-trimethoxytetrahydro-2H-pyran.
 29. The reaction ofclaim 28, further comprising reacting the(R)-3,4,4-trimethoxytetrahydro-2H-pyran with acid to provide(R)-3-methoxydihydro-2H-pyran-4(3H)-one.
 30. The reaction of claim 29wherein the acid is concentrated hydrochloric acid.
 31. The reaction ofclaim 22 wherein the reaction is run at a temperature range from 20° C.to 60° C.
 32. A process for making(R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol comprising

reacting 4,4-dimethoxydihydro-2H-pyran-3(4H)-one with a boron reducingagent and S-(−)-2-methyl-CBS-oxazaborolidine, over a period of at leastsix hours to provide the (R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol in atleast 60% enantiomeric excess.
 33. The process of claim 32 wherein(R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol is formed in at least 90%enantiomeric excess.
 34. The process of claim 32 wherein the boranereducing complex is selected from borane-dimethylsulfide complex orborane-N,N-diethylaniline complex.
 35. The process of claim 34 wherein aTHF solution of the borane-reducing complex andS-(−)-2-methyl-CBS-oxazaborolidine is added to a solution of the4-dimethoxydihydro-2H-pyran-3(4H)-one in THF.
 36. The process of claim32 wherein the reaction is carried out in an inert environment.
 37. Theprocess of claim 36 wherein the reaction is run under nitrogen gas. 38.The reaction of claim 32 further comprising reacting the(R)-4,4-dimethoxytetrahydro-2H-pyran-3-ol with dimethyl sulfate toprovide (S)-3,4,4-trimethoxytetrahydro-2H-pyran.
 39. The reaction ofclaim 38, further comprising reacting the(S)-3,4,4-trimethoxytetrahydro-2H-pyran with acid to provide(S)-3-methoxydihydro-2H-pyran-4(3H)-one.
 40. The reaction of claim 39wherein the acid is concentrated hydrochloric acid.
 41. The reaction ofclaim 32 wherein the reaction is run at a temperature range from 20° C.to 60° C.